Thank you Shiana for an interesting presentation today at journal club. Sorry I did not post a specific place for the 3rd week blog. Thanks to all who posted independently. Please put your 4th week blog here.
Hi guys! Today I had a chance to redo the protein determination procedures that I had messed up last week, and I made sure to be very, very careful with all of the protein samples. I double and triple checked my steps because I really didn't want to mess up again! I also made sure to ask Gladys any questions that I had about the amounts of protein versus water to put in the test tubes. Once I had finished transferring the protein samples into the cuvets for analysis, I became really nervous....because now was the moment of truth!!!!
Gladys came with me to the spectrophotometer because I wanted her to guide me through all of the steps. I told her what I was about to do, and she either said, "Yes, that's correct," or showed me how to do the procedures correctly. After we had run all of the samples through the machine, I saw that my results had come out beautifully!!!!
You can only imagine how happy I was--I had finally gotten through all of the protein determination procedures without any glitches. This time, instead of wanting to yell, "NOOOO!!!!!" I wanted to do a little happy dance and high-five somebody. It was a glorious moment.
I am so glad that I had the chance to fix my previous mistakes; it was a long and tedious process, but I came out with the best result.
Hola everyone! This week is just very interesting. I have been working with UROtsa cells (as usual) and growing and feeding and splitting them. It is all really coming together. I also have been helping Sarah with isolating cells. So isolating cells really involves scraping cells and then using liquid nitrogen to freeze them. It is fun but the liquid nitrogen is not. So, my week was filled with a lot of repetitive stuff like feeding cells and then splitting them. My confidence in the lab is growing, even though it may not be at a rapid pace. I am understanding more about lab equipment and doing a lot of the assays myself. SO my days here are numbered but I am making the most of it.
I also walked to Tiffany and Shiana's lab and found that the rats had come...bad times for me. I am so terrified. Journal club was very informative Shiana! Thanks for that wonderful presentation. Dr. Climecki is also a very good presenter and it really was fun.
Lastly, LBD. He is the boyfriend of Mon-Ning. I know made it happen...*sniffles, cry* But he has been sitting by us lately except for one day...and that got him friend probation. So there is much to do with so little time. I have so many results of western blots (where we examine a certain protein). So I can't wait to make my poster!
Wow. Well, there's pretty much a lot to tell about the 4th week of the internship, but it's all in my lab notebook so there's nothing really to worry about.
This week was mainly focused on trying to get some of the paperwork and the poster done. Well, at least the format done. I'll be borrowing their InDesign program or probably using something. I have to get my paper done, by the way, so I can just paste it on my poster. There really isn't any procedures planned except some explants and PCRs done so that we can get some some of the physiology bit and then Dr. Camenisch and I will plan some sort of one-on-one appointment so that I can get some work done on the abstract. There's so much to do and so little time.
Anyway, I think we had to do a PCR because there was some sort of modification that Derrick wanted to do, while Mark is away. Mark's currently in Washington D.C. Yes he's a very lucky person.
So this week didn't go so well. Demetre and I were almost done with the PCR(ing) but we made even more errors.
Uh-oh #1 I mixed up the Victor Master Mix with the PCR water and ruined the entire stock solution of VMM. That forced us to spend time making more so that we could continue.
Uh-oh #2 Then after finishing 64 samples we realized that in our frantic rush to finish that day we had improperly labeled them. Those had to be thrown out and re-done the next day.
Uh-oh#3 Once we set the samples up for a third time the thermocycler quit on us during cycle 4 once again destroying what we had done.
In Short. We are still not done. However, we are still optimistic. We still have plenty of DNA to keep setting up the PCR's and the only thing we are starting to run out of is time.
-Side Note- On either Thursday or Friday Demetre will be taking me and Daniel out into the field to collect samples! I can't wait!
Review of the past week Last week in the lab was full of laughter! My mentor Tony fired me and rehired me all in the same minute! The reason for this was due to the fact I late to the lab (Less than 5 minutes lol). The night before I had left my cell phone at apple bees and was going crazy looking for it! So then I guess the people at apple bees found my phone and looked through my contact list and called my house early that next morning. It is great to know there are some honest people out there in this world. So I had to find a ride to get to apple bees then manage to get to the lab on time! Fortunately, I have a very understanding mentor and all was good and comical!
In addition, to that moment of comedy in the lab, my lab partner Greg kept instigating little disputes such as how to do PCR and Extractions correctly. More times then not I was correct! Ha ha. Indeed, I have another funny moment in the lab which was when our other lab mentor Demetre broke not one but TWO gels! It was funny because the gels he dropped needed to be disposed of anyways but he thought we actually needed them to run gel electrophoresis. He was fired and rehired as well! Anyways, having this laughter present in the lab allowed for things to run more smoothly as we would all joke around and have a good time while maintaining the serious attitude needed to process lab work correctly.
So to conclude this very meaningful blog I will end with one of my world renowned quotes “Carpe Diem” Seize the day and live life to the fullest.
Helloooo everybody! SO my overall task this past week was trying to get the inesert I hade cloned and replicated in a TA vector in some competent cells out of where they were and into another vector where it could be expressed and measured in S2 cells, and maybe eventually actual mosquitoes. I began with lots of colony PCR, and a digestion with EcoRI on the colonies that worked in order to confirm that my insert was rpesent in the vectors. After the rpesecne of the insert was confirmed, I began digestions with different enzymes (depending on whether it was dealing with the elongation factor gene or 5G1 gene) so I could later directionally ligate my insert into my expression pGL3 vector. I performed overnight digestion on Monday, and was excited to get goin on some progress tuesday.
Tuesday was full of digestions. (I had a biiiig lunch (-: ) But seriously, I digested about 6 different samples, some of which were to isolate a gene of interest and ther were to prepare a vector for the insert to be ligated into. All of my digestions were successful and I ended the day by ligating into my vectors and otssing my DNA into the 14-16 degree overnight.
Wednesday came along and it was the time to make some cells transform (who is excited for TRANSFORMERS THE MOVIE??? MEEEE. But anyways, I transformed and plated some cells to grow overnight. I helped perform a luciferase assay with B then headed over to journal club which was extra informative, for we heard from Shiana, a geneticist ( I <3 genetics..), and learned how to operate InDesign.
thursday was a very sad day. I discovered that my cells had not replicated the DNA I wanted... this meant that either ligations hadn't gone correctly or there was so little ligated DNA that not enough of the cells transformed with it and survived for me to find them. SO i started again, with digestions etc.
Friday was the busiest day I've ever had in my lab, for I got all the way up to the point of ligations over the weekend. That means today I transformed and am ready to walk into the lab and see my wonderful exciting results. (-:
Last week was my journal club presentation on my research article titled "Peroxiredoxin III, a Mitochondrion-specific Peroxidase, which regulates Apoptotic Signaling by Mitochondria." Personally, it took me a long time to understand what the mere title meant. The abstract itself, which I had learned from Katy would give me a general overview of the entire article, became my focus to undecipher for nearly a week before I deigned myself able to move on to the rest. Hence, I was very anxious about being able to condense all of the information I had learned into comprehensible, concise parts for my presentation.
In order to do this, I referenced not only one research article for tidbits of background information and terminology, but many. I recalled the questions that I had when I first began learning about my topic and tried to answer those same queries as they fit into the big picture of what my research will ultimately benefit and contribute to. When the time came for me to present, I was nervous about being able to talk professionally and clearly. Thankfully, my confidence that has steadily been increasing came through for me and I was pleased with how I was able to share my new knowledge. More than one of my peers said that they understood what I tried to impart to them, which made me smile!
Within my lab, I have been able to create my own schedule oriented around my project, which gives me a nice amount of freedom. I am able to let myself into my lab in the mornings and start right away working, though I am often the first person there. Last Sunday, I also came into my lab to carry out a necessary step for the next day—I wanted to save time and make the most of it. I now know where everything I need is and how to use machines like the autoclave and the PCR machine. I have finished constructing two different control vectors by now and am waiting on a sequencing of my mutagenesis primers to come in before I can move on. Prior to last week, mutagenesis did not seem to work, but by tweaking PCR conditions, my mentor was able to fix it at last. This was very exciting news for me because it means that I will be able to move onto the next step soon.
Also! Two-month-old albino rats were received last week and I have been able to help handle them (so that they can get used to humans working with them). They are very cute, but nonetheless I am looking forward to seeing the brain surgery that will be performed on them hopefully at the end of this week. My horizons are most definitely broadening and I love it.
It is hard to believe we all have only less than two weeks left! I am not ready to leave all my new friends or my lab (though with any luck I’ll possibly even be able to stay).
8 comments:
Hi guys!
Today I had a chance to redo the protein determination procedures that I had messed up last week, and I made sure to be very, very careful with all of the protein samples. I double and triple checked my steps because I really didn't want to mess up again! I also made sure to ask Gladys any questions that I had about the amounts of protein versus water to put in the test tubes. Once I had finished transferring the protein samples into the cuvets for analysis, I became really nervous....because now was the moment of truth!!!!
Gladys came with me to the spectrophotometer because I wanted her to guide me through all of the steps. I told her what I was about to do, and she either said, "Yes, that's correct," or showed me how to do the procedures correctly. After we had run all of the samples through the machine, I saw that my results had come out beautifully!!!!
You can only imagine how happy I was--I had finally gotten through all of the protein determination procedures without any glitches. This time, instead of wanting to yell, "NOOOO!!!!!" I wanted to do a little happy dance and high-five somebody. It was a glorious moment.
I am so glad that I had the chance to fix my previous mistakes; it was a long and tedious process, but I came out with the best result.
Hola everyone!
This week is just very interesting. I have been working with UROtsa cells (as usual) and growing and feeding and splitting them. It is all really coming together. I also have been helping Sarah with isolating cells. So isolating cells really involves scraping cells and then using liquid nitrogen to freeze them. It is fun but the liquid nitrogen is not. So, my week was filled with a lot of repetitive stuff like feeding cells and then splitting them. My confidence in the lab is growing, even though it may not be at a rapid pace. I am understanding more about lab equipment and doing a lot of the assays myself. SO my days here are numbered but I am making the most of it.
I also walked to Tiffany and Shiana's lab and found that the rats had come...bad times for me. I am so terrified. Journal club was very informative Shiana! Thanks for that wonderful presentation. Dr. Climecki is also a very good presenter and it really was fun.
Lastly, LBD. He is the boyfriend of Mon-Ning. I know made it happen...*sniffles, cry* But he has been sitting by us lately except for one day...and that got him friend probation. So there is much to do with so little time. I have so many results of western blots (where we examine a certain protein). So I can't wait to make my poster!
Wow. Well, there's pretty much a lot to tell about the 4th week of the internship, but it's all in my lab notebook so there's nothing really to worry about.
This week was mainly focused on trying to get some of the paperwork and the poster done. Well, at least the format done. I'll be borrowing their InDesign program or probably using something. I have to get my paper done, by the way, so I can just paste it on my poster. There really isn't any procedures planned except some explants and PCRs done so that we can get some some of the physiology bit and then Dr. Camenisch and I will plan some sort of one-on-one appointment so that I can get some work done on the abstract. There's so much to do and so little time.
Anyway, I think we had to do a PCR because there was some sort of modification that Derrick wanted to do, while Mark is away. Mark's currently in Washington D.C. Yes he's a very lucky person.
So this week didn't go so well. Demetre and I were almost done with the PCR(ing) but we made even more errors.
Uh-oh #1
I mixed up the Victor Master Mix with the PCR water and ruined the entire stock solution of VMM. That forced us to spend time making more so that we could continue.
Uh-oh #2
Then after finishing 64 samples we realized that in our frantic rush to finish that day we had improperly labeled them. Those had to be thrown out and re-done the next day.
Uh-oh#3
Once we set the samples up for a third time the thermocycler quit on us during cycle 4 once again destroying what we had done.
In Short.
We are still not done. However, we are still optimistic. We still have plenty of DNA to keep setting up the PCR's and the only thing we are starting to run out of is time.
-Side Note- On either Thursday or Friday Demetre will be taking me and Daniel out into the field to collect samples! I can't wait!
Hello Everyone,
Review of the past week Last week in the lab was full of laughter! My mentor Tony fired me and rehired me all in the same minute! The reason for this was due to the fact I late to the lab (Less than 5 minutes lol). The night before I had left my cell phone at apple bees and was going crazy looking for it! So then I guess the people at apple bees found my phone and looked through my contact list and called my house early that next morning. It is great to know there are some honest people out there in this world. So I had to find a ride to get to apple bees then manage to get to the lab on time! Fortunately, I have a very understanding mentor and all was good and comical!
In addition, to that moment of comedy in the lab, my lab partner Greg kept instigating little disputes such as how to do PCR and Extractions correctly. More times then not I was correct! Ha ha. Indeed, I have another funny moment in the lab which was when our other lab mentor Demetre broke not one but TWO gels! It was funny because the gels he dropped needed to be disposed of anyways but he thought we actually needed them to run gel electrophoresis. He was fired and rehired as well! Anyways, having this laughter present in the lab allowed for things to run more smoothly as we would all joke around and have a good time while maintaining the serious attitude needed to process lab work correctly.
So to conclude this very meaningful blog I will end with one of my world renowned quotes “Carpe Diem” Seize the day and live life to the fullest.
Talk to you later amigos!
-Daniel
Helloooo everybody!
SO my overall task this past week was trying to get the inesert I hade cloned and replicated in a TA vector in some competent cells out of where they were and into another vector where it could be expressed and measured in S2 cells, and maybe eventually actual mosquitoes. I began with lots of colony PCR, and a digestion with EcoRI on the colonies that worked in order to confirm that my insert was rpesent in the vectors. After the rpesecne of the insert was confirmed, I began digestions with different enzymes (depending on whether it was dealing with the elongation factor gene or 5G1 gene) so I could later directionally ligate my insert into my expression pGL3 vector. I performed overnight digestion on Monday, and was excited to get goin on some progress tuesday.
Tuesday was full of digestions. (I had a biiiig lunch (-: ) But seriously, I digested about 6 different samples, some of which were to isolate a gene of interest and ther were to prepare a vector for the insert to be ligated into. All of my digestions were successful and I ended the day by ligating into my vectors and otssing my DNA into the 14-16 degree overnight.
Wednesday came along and it was the time to make some cells transform (who is excited for TRANSFORMERS THE MOVIE??? MEEEE. But anyways, I transformed and plated some cells to grow overnight. I helped perform a luciferase assay with B then headed over to journal club which was extra informative, for we heard from Shiana, a geneticist ( I <3 genetics..), and learned how to operate InDesign.
thursday was a very sad day. I discovered that my cells had not replicated the DNA I wanted... this meant that either ligations hadn't gone correctly or there was so little ligated DNA that not enough of the cells transformed with it and survived for me to find them. SO i started again, with digestions etc.
Friday was the busiest day I've ever had in my lab, for I got all the way up to the point of ligations over the weekend. That means today I transformed and am ready to walk into the lab and see my wonderful exciting results. (-:
Last week was my journal club presentation on my research article titled "Peroxiredoxin III, a Mitochondrion-specific Peroxidase, which regulates Apoptotic Signaling by Mitochondria." Personally, it took me a long time to understand what the mere title meant. The abstract itself, which I had learned from Katy would give me a general overview of the entire article, became my focus to undecipher for nearly a week before I deigned myself able to move on to the rest. Hence, I was very anxious about being able to condense all of the information I had learned into comprehensible, concise parts for my presentation.
In order to do this, I referenced not only one research article for tidbits of background information and terminology, but many. I recalled the questions that I had when I first began learning about my topic and tried to answer those same queries as they fit into the big picture of what my research will ultimately benefit and contribute to. When the time came for me to present, I was nervous about being able to talk professionally and clearly. Thankfully, my confidence that has steadily been increasing came through for me and I was pleased with how I was able to share my new knowledge. More than one of my peers said that they understood what I tried to impart to them, which made me smile!
Within my lab, I have been able to create my own schedule oriented around my project, which gives me a nice amount of freedom. I am able to let myself into my lab in the mornings and start right away working, though I am often the first person there. Last Sunday, I also came into my lab to carry out a necessary step for the next day—I wanted to save time and make the most of it. I now know where everything I need is and how to use machines like the autoclave and the PCR machine. I have finished constructing two different control vectors by now and am waiting on a sequencing of my mutagenesis primers to come in before I can move on. Prior to last week, mutagenesis did not seem to work, but by tweaking PCR conditions, my mentor was able to fix it at last. This was very exciting news for me because it means that I will be able to move onto the next step soon.
Also! Two-month-old albino rats were received last week and I have been able to help handle them (so that they can get used to humans working with them). They are very cute, but nonetheless I am looking forward to seeing the brain surgery that will be performed on them hopefully at the end of this week. My horizons are most definitely broadening and I love it.
It is hard to believe we all have only less than two weeks left! I am not ready to leave all my new friends or my lab (though with any luck I’ll possibly even be able to stay).
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