Last Week at CERTL
This week was very easy for me since I had already finished my experiments. I wrote my scripts out and mad sure my powerpoint was neat and good. I also celebrated Joy's birthday with my lab. I just hope that the symposium will be a success for everyone and that everything works out. I really liked this program and hope I can do it again.
A BIG THANKS TO THE CERTL PEOPLE FOR HELPING ME OUT, THE GCRC LECTURERS AND ANDY BREWER FOR HELPING US WITH THE COMPUTER WORKSHOPS !!!!!!!!!!!
Eric
Complete journal:
7-24-06
My last experiments were finished today, the RT-PCR on Orlistat and the Western Blot on Actin. I met up with Steve who told me what to say for my presentations and also revised my PowerPoint a little to fit in with the others. WE have our mock presentations tomorrow and I just want to be prepared so I can do well on it. My RT-PCR turned out pretty good, with good controls and bands. I think that it will be okay to present on Thursday and I hope that my presentation is going to be very good.
7-25-06
Today I went over my script with Steve again and we came to a conclusion on how it would finally look. I am happy with all the work that I managed to get done over these six weeks. I then went to the mock presentations and was what the others did for their projects. I did not get to present however, because Adrianne had to leave at 12. We had our own catered lunch today and it was pretty good. The other’s presentations were also very enlightening. I just hope that the presentations will be good on Thursday.
7-26-06
Today I didn’t do much. I practiced my script a couple of times and then went to my lab to go check up. Since I had finished everything I had to do, I had some free time to chill and hang around. Today was also Joy’s birthday, so we decided to go out for lunch. We went to Botta Bing pizza parlor and had some pizza. Despite it having shrimp and mussels, I still liked it a lot. Joy even got a birthday cake and a candle that sang happy birthday. I had a great time today and I hope that the presentations are going to be good.
7-27-06
Today I practiced my lines some more and got ready for the symposium. Steve went to California so I did not have to go in today for work. I also had to go the mock presentations today since we were cut short on Tuesday. I really enjoyed being in the Kridel lab this summer and I had a great time at CERTL. I hope that the symposium will also be good today. I have learned a great deal from this summer and I hope that I can have to chance to try it again.
5th week at CERTL
This week was the final week that I had to do experiments in my lab. I was worried at first that I would get bogged down with 2 whole experiments to conduct and that I would never finish in time. But thankfully I got some experiments done and I just hope that the outcomes will be good for me. I just look forward to the symposium next week.
Eric
My complete journal:
7-17-06
Today I ran my RT-PCR for XBP1 again and I hope that it would be the last time because I have already done it 3 times before. I also used the proteins that I collected that were treated with Orlistat and ran a gel with them. I hope to transfer them onto a membrane and then put antibodies on them for another western. Time is now a factor for me and I need to get my powerpoint presentation as well as my experiments done in a short period of time. I hope that everything will work out for me as planned and that I do not mess up anymore because that will cost me greatly.
7-18-06
Today I had to put inhibitors on my western blot and then run my RT-PCR. The morning was very busy for me and now my day gets even busier because I also have to treat my cells with Orlistat. I can already tell that my time will have to budgeted because I only have one more week left. This week I also have to do my powerpoint and I think that I will barely be able to get it all done. I ran my RT-PCR for GADD34 and I will conduct an electrophoresis tomorrow, as well as the rest of my western. I went into the dark room to look at my Western, and hope to get good results tomorrow as well.
7-19-06
Today is one of the last days to get experiments done so I took my time wisely and set myself off early in the morning to complete my blot. I also ran my RT-PCR for GADD34 today. I hope that it will work out because tomorrow is my last day to do work and I need every minute of the time to complete an RT-PCR that I do. I also have to put antibodies on my western blot and take a picture of that today. Steve is checking out my PowerPoint to see if it is okay with him so I can present it on Thursday. This week is a week of cramming because experiments need to get done and I should step up the plate if I am to do them correctly.
7-20-06
Today I had to do one final experiment for my RT-PCR. I isolated RNA from my Orlistat treated cells and conducted an RT-PCR on it. I had time to cycle it but I didnÃ’t have time to run it as a gel. In our lab meeting I showed my implications on this 6 weeks and Steve helped me revise my Powerpoint. I was very glad that my final RT-PCR was over because it would be the last experiment I did, and I wouldnÃ’t have to worry so much anymore about getting experiments done. I look forward to presenting my findings to my fellow CERTL students next week.
7-17-06
Today I ran my RT-PCR for XBP1 again and I hope that it would be the last time because I have already done it 3 times before. I also used the proteins that I collected that were treated with Orlistat and ran a gel with them. I hope to transfer them onto a membrane and then put antibodies on them for another western. Time is now a factor for me and I need to get my powerpoint presentation as well as my experiments done in a short period of time. I hope that everything will work out for me as planned and that I do not mess up anymore because that will cost me greatly.
7-18-06
Today I had to put inhibitors on my western blot and then run my RT-PCR. The morning was very busy for me and now my day gets even busier because I also have to treat my cells with Orlistat. I can already tell that my time will have to budgeted because I only have one more week left. This week I also have to do my powerpoint and I think that I will barely be able to get it all done. I ran my RT-PCR for GADD34 and I will conduct an electrophoresis tomorrow, as well as the rest of my western. I went into the dark room to look at my Western, and hope to get good results tomorrow as well.
7-19-06
Today is one of the last days to get experiments done so I took my time wisely and set myself off early in the morning to complete my blot. I also ran my RT-PCR for GADD34 today. I hope that it will work out because tomorrow is my last day to do work and I need every minute of the time to complete an RT-PCR that I do. I also have to put antibodies on my western blot and take a picture of that today. Steve is checking out my PowerPoint to see if it is okay with him so I can present it on Thursday. This week is a week of cramming because experiments need to get done and I should step up the plate if I am to do them correctly.
7-20-06
Today I had to do one final experiment for my RT-PCR. I isolated RNA from my Orlistat treated cells and conducted an RT-PCR on it. I had time to cycle it but I didnÃ’t have time to run it as a gel. In our lab meeting I showed my implications on this 6 weeks and Steve helped me revise my Powerpoint. I was very glad that my final RT-PCR was over because it would be the last experiment I did, and I wouldnÃ’t have to worry so much anymore about getting experiments done. I look forward to presenting my findings to my fellow CERTL students next week.
Fourth week at CERTL
This week I finally got the hang of things around the lab, I think. I did my very own Western Blot this week and I feel proud of myself because it actually worked out for me. I did something right. I still have troubles with RT-PCRs but I'm just glad this week is overEricMy journal this week:7-10-06
Today I ran my gel that I did last Thursday for the protein assay and the lysate collections I had conducted the previous week. I learned that the gel was not enough to read for a western blot and that I needed to transfer the gel into a membrane to be read and photographed. I transferred the gel onto a membrane as a “sandwich” with was composed of a sponge, filter paper, membrane, gel, filter paper, and sponge. I ran an electric current through this whole sandwich to transfer the gel onto the membrane. I also split cells for my next experiments, an RT-PCR on the same thapsigargin treatments, and another Western for the drug Orlistat.
7-11-06
Today I washed my membrane with TBST and put on antibodies for doing the Blot. These antibodies help the membrane bring out the phosphorylation of eIF2alpha on a picture that I would take later. I went inside the dark room and took pictures of my blot. It turned out very nice and similar to ones that have already been done. I have to show total phosphorylation of eIF2alpha on my blot now so that it can be compared to the other blot as a control. I also did my RT-PCR for XBP1 with a DNAse so that I will not get anymore DNA contamination. My cells are not ready to treat so I have to wait until Thursday to treat my cells.
7-12-06
Today I treated my cells with thapsigargin for the RT-PCR that I will do tomorrow. I collected the cells in Trizol since I wanted the RNA from the cells. This time I will be looking for the presence of GADD34 the same time as the phosphorylation of eIF2alpha. I also ran my RT-PCR from yesterday when I had done the XBP1 experiment with the DNAse. This time I hope to get everything perfect and show that I can do a good RT-PCR. I also washed my membrane with total phosphorylation of eIF2alpha to show a control group for my Western experiment.
7-13-06
Today I did another RT-PCR for XBP1 (when are they going to end?) because the last one looked funny (again). So I did every step according to my protocol and will hopefully get it finally right this time. I also isolated RNA from the cells of thapsigargin I collected in Trizol yesterday. I did a totally good western blot though and now I just need to show the same correlation on my RT-PCR for GADD34. I just hope that I do some good work next week and that my gels look good for the western and the other RT-PCR that I will conduct next week.
3rd week at CERTL
This week was better for me because I finally got the hang of doing some of this lab work. I think I finally conducted my first "successful experiment" this week with my XBP1 experiment. I am ready for next week and my new experiments with GADD34.
Eric
Here is my complete journal:
7-3-06
Today was a busy day for me. I had to do a RT-PCR and so that experiment took up all of my day. I also seeded cells for my next experiment on Wednesday. I would be working with the expression of GADD34 as my next experiment. I ran my RT-PCR with Actin loading controls, and XBP1. The experiment would be a success if I see the splicing of the genes in XBP1, but I cannot see this yet for I have to wait until Wednesday to run my gel. The last time I did this I messed up pretty badly, but this time I hope that my mistakes will be fewer and less problematic. I hope that this time around, my RT-PCR will be successful.
7-4-06
Independence Day – NO WORK!!!
7-5-06
Today I had one of the busiest mornings ever. I had two experiments going on at the same time and so I was pretty much occupied until 2 PM. I had to treat and collect my cells for my time course treatment of GADD34. I treated them with 1 uM of Thapsigargin for 30 min, 1hr, 2hrs, 4hrs, and 6hrs. I had to collect my cells at each time point and obtain the proteins from them. I also ran my gel for the RT-PCR that I was doing and got my results. I was pretty happy that the XBP1 in my experiment spliced because that was an indicator that I had done it right. However, my Blanks weren’t very blank, because they had been contaminated with DNA. Still, the results were pretty good for me because it showed that I could do something right (finally).
7-6-06
Today I conducted a protein assay on my GADD34 experiment. This time the assay went quicker and easier than the other times I did it. I thought I did well on it but I ran a scan to see the protein concentrations anyways. I then made my gel for the western blot that I was about to conduct. I poured my gel along with my proteins and mad a membrane for the blot. I would have to leave it over the weekend and then run antibodies over it on Monday. I also ran my gel for the RT-PCR I did yesterday again, but just for the blanks but with a DNAse so there is no contamination on the blanks. I look forward to working out my GADD34 experiment next week.
Second Week at CERTL
The second week at CERTL was a little more settled down, I started the week with my very first experiment, and had a wonderful time doing it. But as time went on in the week, I had more and more time to do things because my cells had to be split and I had to wait a day until they could be used. I had some down time for a change, and I liked it. All in all, this week has been pretty good for me.
Eric
Here is my full journal on the week:
6-26-06
I worked on my first official experiment today. I am now conducting a research project on the processing of XBP-1 and I had to retrieve and isolate RNA from my cells in order to do this. I had to come in on Friday to split and seed my cells, because they would have died out if I had not. I treated one of my experimental groups today with a drug called thapsigargin, and conducted a RT-PCR with the cells I obtained. After I isolated the RNA, I had to make them into solutions with a positive and negative control in order for the experiment to be controlled. I obtained cDNA from the RNA and used it in a PCR and am now currently preparing to run a gel tomorrow.
6-27-06
Today I ran the gel for my PCR and found out the results. Although they were not what I had expected, I did manage to do part of it right. The results were a little funky with the loading controls because they contained DNA bands (which aren’t supposed to be there). I did get my negative control, thapsigargin, to splice, which is supposed to happen in this experiment. I was treating to show ER stress by the splicing of a gene called XBP-1, which did splice in my thapsigargin control, and also a little in C75, an anti tumor drug used to inhibit FAS. I also cultured some more cells for another RT-PCR that I am doing next week.
6-28-06
Today I didn’t do much lab work, because my cells were just split and haven’t grown enough for me to isolate the RNA from them. I just sort of sat back and chilled for a bit today. I found out from Steve that I will be performing a western blot along with a RT-PCR on my cells in order to find a specific gene called GADD34. I will be researching the presence of it when treated with FAS inhibitors. The inhibitors induce ER stress, and it causes apoptosis of the cell with the phosphorylation of eIF2alpha. GADD34 comes into play with the phosphorylation of eIF2alpha, and it dephosphorylizes the eIF2alpha, in order to retain homeostasis. I will find out it the cells induce GADD34 with treatments. I also went in the dark room with Joy and made pictures of her western blot.
6-29-06
Today was also not a busy day for me. I didn’t have anything to do for the morning, because treating my cells in the morning meant that I would have to come in tomorrow morning and collect, which is impossible for me to do. So, I treated my cells with DMSO, Orlistat, C75, and thapsigargin in the afternoon. I have to come in tomorrow afternoon to freeze the cells in Trizol, so they can be kept in storage over the weekend. I treated my cells by mixing media with the FAS inhibitors and then applying it to my cells. These cells would be left with the drugs for a specific amount of time if I was doing a time course test, but I wasn’t, and so I just treated all of them with different drugs.
First Week at CERTL Summer Symposium
This is the first week at the CERTL Program
The First week was:
The first week was pretty cool, but otherwise very tiring for me. I was overwhelmed with different kinds of stuff and new science lingo that I had never heard of before. I got to do hands on stuff with cancer cells, and learned how to do many methods of obtaining different results. I learned how to culture cells and seed them into different plates to keep them living. I learned that you have to add media, or their source of food, for them to grow. There are also different ways to obtain "information" from them by utilizing different experiments. I learned how to conduct an RT-PCR, which uses the RNA from the cells. I had to give a speech on the RT-PCR process to my mentor. Here is the PCR process:
I also did a protein assay to analyze the protein concentrations in my cells. I messed up 2 times before I finally got it right. I then got to run in on a gel and am currently conducting a western blot. I just prepared the gel "sandwich" on Thursday. All in all, this week has been alright for me, because the experiments were cool, but they got to be so overwhelming for me.
Eric
Here is a detailed description of my week:
6-19-06
Today I came in for the first time as a scientist in Dr. Kridel’s lab. His graduate student, Joy, is going to be teaching me how to do some experiments this week, such as a Western Blot or how to isolate RNA. I learned how to seed cells today, and some scientific lingo such as to aspirate, or to suck up (literally). I also learned how to mix media, or the food used by cells. I got to have hands on experience in seeding new cancer cells, by first cleaning them with a saline solution, and the using a pipette to mix in media with the cells.
6-20-06
Today we learned how to isolate RNA from cells. I learned that we used Trizol and Chloroform to first let out the RNA from the cells, and then we used a centrifuge to isolate only the RNA for testing. After this we got to calculate the amount of RNA in our solution, and we used a spectrophotometer to analyze RNA concentrations. We added some RT enzymes to the solution and Reverse transcribed it into cDNA. We then ran a PCR on the DNA and found out how it looked.
6-21-06
Today I learned how to do a protein assay. We took protein from cells and set them up for the computer to scan and see how much protein the cells had. We also ran a gel electrophoresis on our DNA samples and discovered the results to our experiment. I put the results into my lab notebook and completed my first official “experiment.” I have to prepare for an informative speech to Dr. Kridel about how RT-PCR works tomorrow. Also we are setting up to do a Western Blot, which we hope to conduct tomorrow morning.
6-22-06
I had to redo the protein assay because it didn’t work out as I had expected. The protein assay takes a long time to do and is very tedious to do, but I managed to complete another one by lunch. This time the assay was successful and so now I could work on the Western Blot. We prepared a gel for the proteins and conducted a Western Blot. The gel has to sit and be run later because there needs to be time for it to dry. I presented my knowledge on RT-PCR for Dr. Kridel today and we figured out an appropriate title for my PowerPoint. The first week in the lab was mostly okay, except for me messing up on the Protein assay. I enjoyed learning about how to conduct the experiments and look forward to next week.
