Week 2: Microbial Ecology Laboratory, Iowa State University

January 21st, 2015

Kelsey King ’15, Bahnick Fellow in Ecology, Evolution & Organismal Biology

This week was the busiest of my entire time, lab-wise. We began the final 60-day incubation study. On Monday, I filled and labeled 63 sample vials. Then I made solutions to be used as the experimental substrate. The downside of classes beginning is that I no longer have the lab all to myself. The undergraduate research assistants are back. On the plus side, I do have a small team of students that can help me wash my dishes–amazing! I still peel labels and remove the peat soil, though. I’ve been there with the uber-dirty dishes, and it’s not fun.

Sample vial

Sample vial

Tuesday was the big day. The day where all the solutions were completed and added to the experimental vials. Then the vials were sealed, and all CO2 was removed from their air. If I messed anything up here, I could ruin the experiment. It’s not too worrisome, though, because I have plenty of other places where I could make an experiment-ending mistake. Needless to say, after this day I was exhausted.

Flushing vials (29/63)

Flushing vials (29/63)

The next day the first step was clean up. All these solutions made a lot of dishes (too many for the lab assistants to do on their own and keep up with all the other stuff happening in the lab). Then I sampled the gasses inside the vials and measured how much CO2 had accumulated since I cleared the air. And then there was plenty of data to be entered into the computer. I’ve learned this at Cornell: never put off entering data and setting up your analysis.

Sampling

Sampling

Morning dishes again on Thursday. However, it was also lab meeting day. We reviewed a paper that is in the process of being published (outside of the lab group, but pertinent to much of the lab’s research). Amazing. Everyone went around and talked both about the good writing that was in this article (which will be published in the Ecological Society of America) and the science. Nothing has been more stimulating than watching the other people in this meeting sharing their expertise, because we all come from different scientific backgrounds, and discussing ingenuity and the flaws of this experiment. The amazing part was that I even got to contribute something useful to the discussion. Being a part of discussions like these are invaluable experiences that just aren’t quite the same as in a group of undergraduates.

Back to my experiment again where I added moisture to our incubating samples because we don’t want the moisture levels to drop beneath a certain percent, since moisture is very important in the bog system when it comes to the function of the decomposers. The peat we’re using features a depth approximately 30cm below the water and up to 20cm above the water. This sample peat therefore has all sorts of decomposers that work best at varying moisture levels from underwater to baking in the sun. Since moisture is not a variable we’re testing, we need it to be consistent.

Friday!  Dishes again. Unfortunately, the lab assistants are not in on Friday mornings and so there is no one to kindly get to my dishes before I have a chance to. Then we reviewed data, looked at the methods I’ve written up, and flushed our samples to prepare for the Saturday sampling. Working on Saturdays is a bit creepy because the building tends to be pretty much empty, but I do love working in the building when it’s quiet. Among other things, I’ve discovered that when people talk to me while I’m working in a lab (as opposed to the field), I tend to mess up.

My work station

My work station

Iowa State University is vastly different from Cornell College.  Instead of the huge, welcoming, close community I feel at Cornell, there are small pockets of community and friendship here. I always knew I loved Cornell and that I’m lucky to have found the school, but I never imagined how much I would’ve disliked being an undergraduate at a large public school. I definitely found the right school for me.

Professionally, however, I do think I would enjoy working at a large school.  There’s so many interesting projects going on all the time, and having access to all the wonderful scientists in our lab and in the same building is really great. I’m going to make a point next week to pay a visit to a few other labs while I’m here, just to see what they’re like and meet more researchers.

 

Week 6 & 7: Teacher Created Materials

January 16th, 2015

James Thompson ’15, Hanson Fellow in Accounting

TCM Last Day

Outside the TCM Building! (Picture taken by Jared)

My last couple of weeks for the fellowship have come to a close. Week 6 was a continuation of holiday break in the office, and I spent my time relaxing and preparing for the year ahead. During the last week of my fellowship, much of what I learned about accounting at TCM came together as I worked with Senior Accountant Jared Silva in the Gains & Losses department.

The most important activities I did in Gains & Losses over the course of the week were to create and automate financial reports and statements. Some of the reports and statements were:

  • Q4 Financial Results. This was the biggest report I got to work with, pulling and combining a lot of data into different groupings. While helping put the report together, I acquired a better understanding of how the business runs on a quarterly and annual basis, rather than on a daily or weekly basis as I had been accustomed to seeing earlier.
  • Operating Report. This is to ensure that the money in transit between the bank and TCM is in balance. It needs to be reconciled monthly, although at some larger companies, this is done weekly or daily. If the report is not reconciled regularly, problems can accumulate and roll onto the next period’s report, adding to the magnitude of issues to resolve. Data has to be drawn from the A/P and A/R departments, so solving issues can require the entire department working together to tell the story behind misplaced numbers.
  • Daily Financial Dashboard. Jared sends out a report every morning, summarizing the company’s position across multiple fronts, including total revenue, costs, and inventory, and compares that to the same month in previous times last year. It makes sense for TCM to make comparisons on a monthly basis since their business is seasonal in nature. I helped update the report for the new year, fixing any bugs Jared noticed and making improvements wherever I saw fit.
  • Miscellaneous. I did not realize how many small reports here and there are requested of Jared. One day Jared will get a request from the marketing division; the next day, the sales division; and the day after that, the CFO. The small reports add up to a considerable amount of extra work. I helped him out in this area. Since there is no set process for creating any of these type of reports, figuring out where to find the data is the trickiest part. We needed the assistance of the IT department on a couple of occasions to find what we needed, and they were very supportive.

One byproduct of automating and creating these reports was becoming better at using formulas in Excel, especially “VLOOKUP.” I already had quite a bit of experience creating formulas in Excel, but every problem requires a different solution, so I ended up learning some new formulas along the way through the great Google. Cornell has provided me with the tools to assimilate information quickly, and that particularly came in handy this week.

Overall, I know that accounting is not the field that I am interested in pursuing, but that is beside the point to how much I have learned from this internship. The knowledge I have gained will be applicable in any finance related career that I go into. On top of that, the technical skills in software such as Excel will be of use across many job types. I am extraordinarily thankful for being given the chance to go to California and do a fellowship with Teacher Created Materials.

Week 1: Microbial Ecology Laboratory, Iowa State University

January 14th, 2015

Kelsey King ’15,  Bahnick Fellow in Ecology, Evolution & Organismal Biology

My internship at Iowa State focuses on bogs. They don’t smell great, but they’re actually pretty great at providing various ecosystem services. The most important one, which could really be disturbed, is their ability to be a carbon sink. With climate change on the rise, the environmental restraints–such as temperature–could be shifted to the point that a particular bog is no longer a carbon sink but a carbon source. SPRUCE is a project that is investigating these possibilities, and the project that I’m now a part of for four and a half weeks.

I’m working with Dr. Kirsten Hofmockel’s lab with one of the researchers there, Dr. Ashley Keiser. This is a lab full of post-docs and one doctoral student. My first week at Iowa State was most importantly lab orientation, and experimental preps. The timing of this week was really wonderful because I got to be involved in the experimental design, and also got used to the new foreign campus without all of the students here. After lab introductions, it was time to begin the preparations for a 60-day incubation that I will set up and begin testing. We began by discussing the best way to utilize various samples to make up our experimental peat. Which ended up entailing hand-mixing mossy soil-like water.

Freshly mixed peat

Freshly mixed peat

This week I’ve learned new methods on testing the properties of soils like pH, or moisture content. I also milled soil, and completed a run through of our experiment to determine the appropriate mass of the peat to be used in the final 60-day incubation. All of the stock solutions have been made, after of course I learned how to use a microbalance, and a large stack of to-read papers have been collected. The most important method I learned this week was operating and loading the machine that analyzes the amount of CO2 in a sample. This is the machine that I will be using to collect all of the data for my experiment, and I’m proud to say that I can use the syringe without shedding blood.

IMG_1776

Moisture test

However, the most enlightening part of the week was when I attended a lab meeting with the doctoral student, the post docs, and Dr. Hofmockel. This meeting showed me not only how the lab is run, but how much the interpersonal component plays in to graduate and post-graduate work. The lab meeting’s topic was a conference involving soil biodiversity, which is perfect for this lab of soil microecology. I got to hear a review of the conference with critiques of the set up, and general information that may be useful to different projects going on in the lab. It was great to see the community structure of the lab, and how much everyone is willing to help each other out.

The most surprising part of my first week was how easy it was to begin fitting in to the lab, and just doing the lab work. I suppose before I came to the lab I was concerned that I didn’t have enough experience doing lab work, or that I’d be there doing the manual labor and not necessarily controlling the research.The partnership that Dr. Keiser and I have is wonderful, and I’m glad to know that Cornell isn’t the only place that will trust undergraduates to do the real work.

Though my Cornell experience hasn’t involved working with soils, it has provided me invaluable lab experience. The amount of experience I’ve had throughout my Cornell courses has allowed me to feel very comfortable doing basic lab procedures like making stock solutions, washing dishes, and even thinking through experimental design problems. This confidence has allowed me to speak up when I have a solution to a problem. Something I don’t think many semester schools really cultivate in their undergraduates.

Week 5: Teacher Created Materials

January 6th, 2015

James Thompson ’15 Hanson Fellow in Accounting

This week was exceptionally short due to the holiday season. I went to work for two days, December 22nd and 23rd, and the office was quiet since some people already took off for the holidays. We officially finished the audit, sent out more letters for A/R to collect on overdue invoices, and organized the filing cabinets for the upcoming year. The office is in tip-top shape since we used the couple of days to wrap up the smaller tasks that get pushed to the back burner during the busier days. As I have mentioned, being organized is paramount during audits. Keeping the office organized is the basis to tracking many documents down, so it needs to be done whenever the time presents itself. With my last week at TCM coming up, I am looking forward to assisting Jared, a senior accountant, in Gains & Losses (G/L) during the year end closing procedure.

I am celebrating the holiday season with my girlfriend, who is home from college, and her family. We stayed in California through Christmas, had a classic, delicious Christmas meal with the extended family, and took off to Utah. The backdrop of the mountains is stunning, although I will say it is easier to enjoy from the comfort of the living room. Board games, target shooting, and socializing with the neighbors have made the holiday break pleasant and relaxing.

Merry Christmas and Happy New Year!

Week 4: Teacher Created Materials

January 6th, 2015

James Thompson ’15 Hanson Fellow in Accounting

This week at Teacher Created Materials has been different from the normal day-to-day accounting department operations. We are going through a semi-annual audit before the end of the year. Personally, I have been doing many different tasks to help the audit go smoother and quicker. By doing so, I have seen the fundamentals of how audits are conducted; I have watched the relationship between the company and the auditor; and I have a better grasp on what it entails to go through an audit. I’m going to sum up this week’s experience through Q&A because that is what my week most resembled. I have never seen an audit being conducted, so watching it happen made me ask many questions and figure out some answers.

What is the purpose of an audit?

An audit is done to check the accuracy of the financial reports. It is conducted by an independent party so that there is no finagling involved. For an inventory-heavy company such as TCM, a critical aspect of the audit is to make sure the inventory is properly maintained and valued.

How is this done?

It would be far too tedious and time consuming to check each and every line item on every report turned out by the accounting office. That’s not a practical approach. Instead, sample testing is used to conduct the audit. The auditor will take samples for different aspects of the company (inventory, credit memos, invoices, etc.). Then, in turn, we have to provide him any information he requests.

Where does the information come from?

We need to provide primary source information. That means we need to give him check stubs, invoices, purchase orders, and proofs of delivery to show that certain transactions did in fact occur. The level of organization within the accounting department comes into light during this. There is a plethora of data to sift through to find the paperwork of a single transaction, so the quality and accuracy of the data input into the accounting computer systems really, really matters. To find a specific document, which could be anywhere inside multiple filing cabinets chock-full of paper, within a matter of minutes seems only a little short of miraculous.

Other things I have learned about audits.

Different auditors have different preferences. Since we have an auditor that is new to working with TCM, the question of how much evidence to supply to the auditor popped up frequently during discussions. Giving the auditor an inappropriate scope of documents could make the audit take far longer and be more demanding than it already is.

Overall, I can see how doing an audit is essential to keeping a company honest, organized, and accurate in its practice. Cornell has taught me how looking at primary documents and piecing them together, such as the manuscripts for the East India Company we looked at during Economics of Organizations, can tell a full story. It takes in-depth research and educated inferences to make the parts fit together. Accounting during audits is the same at its core.

 

 

Week 3: Teacher Created Materials

December 16th, 2014

James Thompson ’15 Hanson Fellow in Accounting

The third week at Teacher Created Materials has been a turning point in my internship. I spent more time working in A/P and had the opportunity to attend a day-long training session for employees to learn about the company.

My time in A/P began to get more in-depth this week. As year end is quickly approaching, Sheila needed as much help as possible to finish her A/P tasks before their deadlines. Some of the tasks I got to spend time on are:

  • Differentiating between selling cost and the cost of products. The difference between these two things is important to account for the profitability of sales representatives. When expenses are placed under selling costs, it means that extra costs were incurred by the sales reps while trying to sell a product, hurting the profitability of sales reps. However, if costs are placed under the cost of products, the costs will not affect sales reps at all. The costs will simply be deducted from the company revenue.
  • Making transfers from assets to expenses. Certain costs from vendors needed to be placed under expenses although they were initially recorded as assets. I made these transfers using SAGE (accounting software) and Excel. SAGE is not intuitive since it requires understanding the coding system the accounting department in TCM uses to record each entry. Taking the code on certain assets and changing them to be listed as an expense is all that needed to be done, but that required using an Excel spreadsheet created by Sheila that is then uploaded into SAGE. It’s a specific, time consuming process that Sheila has learned to streamline using Excel.
  • Expensewire. TCM reimburses daycare fees for any employees with children younger than a determined age.  Sheila has created an Excel spreadsheet to speed up the necessary calculations. There was some manual entry that she wanted to eliminate since it was redundant, so she assigned me the task of figuring out how to automate the spreadsheet. After an hour of trying to make the spreadsheet do what I wanted it to, I realized the solution requires programming a Macro, and I don’t know how to do that. So we called in one a person from IT, and they were able to do the programming for us! Now I really want to learn more about coding. After that was solved, we were able to upload the expenses into SAGE faster than before.
  • Used an Amortization Schedule for journal entries. If not for financial accounting, I would not have known how to interpret an amortization schedule. Using the date of an invoice listed on the amortization schedule, I had to account for part of the payment balance under principle and the rest under interest. It’s akin to a mortgage.
  • Reconciliation. I reconciled a few credit card statements with the charges issued to TCM. Prof. Santhi Heejebu taught me how to use Pivot Tables during Business Analytics, and that came in handy during reconciliation! Pivot Tables make doing a reconciliation much easier and faster.

I also learned more about the company during TCM University. It is a day long event to help employees understand the big picture of TCM. At its essence, TCM strives for “joy in learning and wonder in learning,” and their products illustrate this approach. I also learned that TCM was started out of a garage in 1977 by a teacher trying to publish her book, Quick Fun Art. After being rejected by multiple publishers, Rachelle Cracchiolo decided to go the entrepreneurial route by publishing for herself! It’s inspiring to consider how far the company has come. In addition to interesting stories, I learned about the various departments in the company, so I have a notion of how the company operates altogether.

TCM University

I’m officially part of the first TCM University graduating class!

At the event, employees were very passionate about working for a company that aligns with their views. Working to create engaging educational materials so that students like to learn was by far the most frequently cited reason people want to work for TCM. It helped me realize that working for a company that has a vision I feel strongly about is valuable, and it is a thought I will keep in mind as I start my career after graduation.

This weekend I got to play more soccer! (Go figure.) I was part of a team that played in the L.A. Holiday Classic Soccer Tournament. It is a 7 vs 7 format and it took place on the training fields right outside the StubHub Center, home to the L.A. Galaxy. We had a cohesive team, and were undefeated in the men’s over 20 division until the championship game. More importantly, it was super fun. As the result of an unbiased decision by my teammates, I was bestowed a banana for being the Golden Boot of the tournament. To celebrate we gorged on tasty Mexican seafood dishes at El Pescador. I think I am still full.

Our team - the OC Lions.

Our team of goofballs – the OC Lions.

 

Week 2: Teacher Created Materials

December 8th, 2014

James Thompson ’15 Hanson Fellow in Accounting

The overwhelming majority of my time during the second week with TCM has been spent working on physical inventory. This year’s physical inventory has been their most intense to date. TCM does their physical inventory manually, using counting, paper, pen, and data entry. They are not quite large enough as a business to warrant an electronic physical inventory system, although that would make the process much less strenuous.

Physical inventory is necessary at year end in order to file taxes and be credible during audits. It’s necessary for every business, especially businesses with lots of inventory. Lisa had me take part in each of the different aspects of physical inventory to help me gain an understanding of a few processes. Here are the steps in physical inventory:

  1. Count the books in the warehouse. Each worker is given paper with a number and a plenty of rows. The number stands for the bin in the warehouse where inventory is taking place. A bin is a specific part of a shelf, and there are plenty of bins. Within each bin the book name, code, and number of books needs to be written in the rows for each different book. This is a time-consuming process, since it requires literally counting all of the books in the warehouse.
  2. Enter the data. The data on each piece of paper needs to be entered into Excel. While entering the data, a VLookup function is used to make sure the name of each book matches up to the code written down on the piece of paper.
  3. Sort the paper. The paper numbers can get jumbled up after going from the warehouse to the accounting office since they exchange hands a few times. Yet, for the papers to be of any use as evidence, we need to be able to reference them. The only way to reference a particular piece of paper is for all the paper to be placed in number order. This posed a daunting task since there is so much paper. In essence, I created a real life sorting algorithm to finish the task.

    All of the paper I sorted into numerical order -- an 18 hour project.

    All of the paper I sorted into numerical order — an 18-hour project.

  4. Double check. Lastly, we need to check for missing pieces of paper (missing data) and check for items the system says we have but we did not count (“Frozen Not Counted”). Either could result in an inaccurate report of the value of TCM’s inventory.

When evaluating the data after the project, a variance test is done, which is basically a simple statistics test. First, the accounting department uses what data we have in the system previous to the physical inventory to estimate how much inventory is probably in each bin. Then, after the physical inventory, any inventory value over/under by a certain amount compared to the estimate is re-evaluated. All-in-all, this is explaining to me how accounting departments keep accurate records of their finances in the midst of so much inventory moving in and out of the business. It is clear that every business needs to use a rigorous system to keep their financial records healthy.

Besides working on physical inventory with TCM, I spent much of the week indoors. I was caught off-guard by the weather! In a rare event, SoCal had a few days worth of rain, causing mudslides in Malibu and forcing me to walk to work rather than ride a bike. It was funny to see what the SoCal version of a “storm” is — quite mild. But I was able to get out and play soccer on some unfamiliar fields on Friday night and Saturday morning! On top of that, I got to go to Huntington Beach Pier on Sunday and soak up the California sun that I expected to see all the time.

Week 1: Teacher Created Materials

December 2nd, 2014

James Thompson ’15, Hanson Fellow in Accounting

Moving to Huntington Beach, CA for this fellowship with Teacher Created Materials has been a completely new social experience for me. There are a couple of apparent differences between living on my own and living at school:

  1. I have to make food for myself. Yikes! I appreciate Bon Appetit even more than I did before.
  2. I lack friends around this part of the world.

There is a grocery store near my house, so the first part does not pose much of an issue. However, I have to go out and meet people in Huntington Beach if I want to meet people outside of work. My second day in the area was a Saturday, the preferred day of the week for soccer. By searching online, I was fortunate to find a pick-up soccer group playing a game for the Children’s Hospital of Orange County. Everyone is welcome, as long as you bring a few dollars for charity. The combination of a great cause and a sport I can’t get enough made meeting people easy, and on top of that, we raised $300 by having fun! I feel more connected to the community, too.

The delightful community of soccer players!

The delightful community of soccer players!

Monday was my first day with Teacher Created Materials (TCM). TCM is a business that creates and sells educational materials for schools and teachers in K-12. I am going to be learning the different parts of accounting during my time with their business. Lisa Whiteman, Controller, laid out how my time with TCM will go. I will be spending time in:

  1. Accounts Receivable (A/R)
  2. Accounts Payable (A/P)
  3. Gains and Losses (G/L)

That is also the order I will be going through the sections, which should keep a steady increase in the complexity of projects and job shadowing that I am doing. In addition, there will be a few miscellaneous days, such as physically accounting for inventory in the warehouse and doing a day-long course for employees of TCM. By the end of my time with TCM, Lisa assures me that I will either “love or hate” accounting, but no matter the case, I will know more than I did.

This is my work space in the accounting office.

This is my workspace in the accounting office.

For the first week at TCM, the office is quieter than usual since Thanksgiving break is right around the corner. Regardless, the accounting team has to keep the business in check. I spent time in A/R with Vanessa working on a project for the entirety of the week.

The project entails trying to increase cash flows by contacting businesses with delinquent payments. The end of the year is coming up, making this a crucial time to receive payments. Basically, I found delinquent invoices (30+ days overdue) and the according proof of delivery (POD), printed those out, and attached a memo of why we are contacting their business. Undertaking this project required me to learn the basics of navigating ELAN, an internal accounting and customer service software TCM uses.  I sent out quite a few notices, so hopefully cash flows increase within the next few weeks.

While working on the delinquent payments project, I learned about how Vanessa got her start with TCM. She began with TCM in a part-time customer service position while going to school full-time. For two years, Vanessa worked hard, showing her dependability and organization. Then, three years ago, she was offered a full-time position as the A/R for TCM. She could not leave the opportunity hanging, but taking on work and school full-time would have been too much, so she now goes to school for accounting part-time while working full-time for TCM. Her determination is remarkable.

Our savory Thanksgiving meal, featuring rare prime rib.

Our savory Thanksgiving meal, featuring rare prime rib.

To finish off the first week-and-a-half I have spent in California, I celebrated Thanksgiving with my girlfriend’s family just north of Santa Monica! Her grandma, uncles, and I dined in excess, competed in board games, and laughed incessantly for the whole break. It was undoubtedly a heart warming get together. I am so fortunate to have her family in SoCal welcome me into their home!

Week 9: Children’s Hospital Colorado

August 7th, 2014

Sandra Cordero ’16, Farley Fellow in Children’s Research 

Although, it is my last week, I been having trouble logging into EPIC and with my badge access. There was a misunderstanding and I was terminated earlier than I was supposed to, thus had to deal with this situation to continue my data collecting. However, this gave me time to work on my PowerPoint presentation, which I received help with. Originally, when I thought that I would not have any results to present, I was going to spend time giving a background on the different surgical treatments that we focused on. Stating the advantages and disadvantages of each treatment, and explain some past studies. However, Erin thought that people will be more interested in learning my study instead of others, and thought that I would spend a lot of time introducing these treatments instead of the literature of my study. Thus, it was a very last minute decision, but I asked Patrick to do a preliminary result summary on the demographics of my study and associated complications. He was not able to do a full statistical analysis, but I was able to learn about the need for physical therapy by groups (SMP, TENs, and IM Nail), the need for re-operation by group, have a mean table for the average age and weight for each group, and surgical rate by group for fracture stability. However, I was still able to include the advantages and disadvantages of each surgical treatment, but I chose to put these slides in the discussion section to tie it back to our overall findings. My discussion was SMP is an effective treatment for unstable femur fractures.

This summer I learned how to accept good criticism. When I did a practice presentation with Patrick and Erin, they helped me re-design my PowerPoint and gave me tips to help my presentation flow better and prepare me for questions that I may be asked. One skill that I learned was how to report results. Originally, I had my tables and graphs by counts, but since we were comparing among groups, I had to change these numbers into percentage, since there was not a same amount of observations for each group. In addition, they helped me with my transitions. For instance I did a short introduction explaining what each surgical treatment was. Patrick recommended that I state (during my presentation) that I will refer back to these treatments in my discussion section and relate it back to my findings. Since I only explained SMP, TENs, rigid intramedullary nailing, and Trochanteric Entry Nailing and not the others, it might be confusing to the audience. They also helped me re-define and re-word my research question and hypothesis. During my presentation, I was quite nervous, but I was proud of myself, I was able to say what I wanted to say and explain these topics effectively. Patrick was impressed that I was also able to answer the follow-up questions, and told me it showed that I was very knowledgeable about my topic and prepared for my presentation.

This internship was an amazing experience. I think that over the course of this summer, I was able to gain more confidence in myself. I was the youngest intern at the CHCO, and it was quite overwhelming sometimes since the rest of the interns were graduated and were in the process of applying to medical school. However, I felt that I handled the situation well. There were times I doubted myself and the reasons I was there. Hence, why I was quiet and reserved the first couple of weeks. But, I think once I got to know my colleagues I learned that I shouldn’t be afraid of them, and that I should look up to them for inspiration and advice for my future.  They were a wonderful group to work with and I have learned a lot from each one of them.

It was a great  summer, being able to work in Colorado and build lasting friendships; especially with other Cornell students, this is something that I will treasure forever. Thank you RJ and the Cornell Fellows program  for this wonderful opportunity.

After I finished my presentation, I received a certificate of completion.

After I finished my presentation, I received a certificate of completion.

 

Week 11: Pufall Lab, University of Iowa

August 7th, 2014

John Christiansen ’15, Dimensions Fellow in Research

This week has been spent trying to get the deletion check primers to work. These PCR primers would be used to amplify a region of transfected NALM6 genomic DNA to determine if the CRISPR vectors were effective in cutting out their target. However, previous attempts to validate the effectiveness of these primers using wild type NAML6 DNA have not gone well. I later learned that the primers were misordered. This week we received the proper primers and I began to work with them.

So far, I have performed the PCR with these primers three times. The first time (Friday, August 1st) I found that none of my samples, even the positive control, had any product bands when analyzed with gel electrophoresis, though the 100 bp ladders still showed up clearly. This suggested that something had been wrong in the reaction, and on closer examination it was found that I had added too much dNTP. PCR can be finicky and this might be why nothing was amplified. On Monday (August 4th) I tried again. This time the correct dNTP concentration was used, but the only product bands seen were two very faint bands, each in no template control lanes. Oddly enough they were the right size for the primers used. The procedure was performed correctly, so the fact that I saw so few bands suggested that my reactants were somehow inept and the fact that I saw bands in the no template control suggested that I had DNA contamination. The solution to both problems was to replace each reagent and try again. I also reduced the amount of ladder I loaded in with my samples after being informed that the imaging machine we use to see the bands generates an image based on relative intensity.

The third time I tried PCR with the correct primers (August 5th) I used new reagents and only loaded 3 uL of the ladder with the samples. I finally got some positive results. Out of the five primer sets tested, two had worked very well. One had replicated the desired product well but had faintly replaced a few other bands, and another produced appropriately sized but faint product bands. Only one of the five sets failed to produce a product band. These results were not perfect, but it was a relief to finally see positive results. Later today (August 6th) I’ll be redoing the three sets that didn’t work as well with some different conditions to see if I can optimize them.

Of the five primer sets, two were for my work with BCL6 and the other three were for different gene that the lab technician Mimi had been working on. My two primers worked out well, and I am ready to move on with deletion screening as soon as Mimi has the time to show me how that procedure works; hopefully I’ll be able to pick it up from watching her work with her primers. I also have assembled the samples I needed to test the dexamethasone induced BCL6 regulation in SUP-B15 cells. I have previously do the same with NALM6 cells, but a few complication have previously delayed my work on SUP-B15. I look forward to retesting the qPCR skills I have developed this summer.

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