So as I explained last week there was a slight change of plans. The non functioning tropomyosin caused a few alterations and I was suddenly a detective on a different case. A case I thought I was going to follow up this week. However, due to science being more expensive and taking more time than I might have I was led on to another trail these week.
As the tropomyosin mutations didn't work my supervisor was kind enough to supply me with some troponin mutations. Troponin is also part of the thin filament so the effects of the mutations would be possible to investigate the same way as the tropomyosin mutations would have been originally. Hence this is what I did. And this time I was provided with results that one could understand. However, this has now provided me with a lot of data for my report. The report I will write next week as it is my LAST one :(
Sunday 15 August 2010
Friday 6 August 2010
We need PROOF!
So dear readers as I’ve mentioned the aim for my 7 weeks here in the lab was to investigate the effects of tropomyosin mutations causing skeletal muscle myopathise. This being done with the aid of ATPase assays. As you might have understood by my earlier blogs, ATPase assays is something I’ve done a lot and without blowing my own horn I would say it’s kind of my specialty now. So imagine my surprise when doing them this week with the mutated tropomyosin and getting weird results indeed. Only doubting my skills slightly I re-did the assay until I was in fear of running out of my precious mutant and still I had no understandable data. Now naturally this is science, things don’t always turn out the way you hoped. But fear not, you always have to joy of investigating WHY it didn’t turn out as expected.
My supervisor was sure the mutant had not been transcribed properly, but one cannot be sure without proof. So proof I got. Well tried to get. Many things were investigated and finally a co-sedimentation showed us the light, the mutant wasn’t binding to actin. Obviously my supervisor had his ideas about why the mutant wasn’t binding to actin. However, proof is still needed for these ideas too, so it’s pretty obvious what I’ll be doing next week isn’t it.
My supervisor was sure the mutant had not been transcribed properly, but one cannot be sure without proof. So proof I got. Well tried to get. Many things were investigated and finally a co-sedimentation showed us the light, the mutant wasn’t binding to actin. Obviously my supervisor had his ideas about why the mutant wasn’t binding to actin. However, proof is still needed for these ideas too, so it’s pretty obvious what I’ll be doing next week isn’t it.
Friday 30 July 2010
Jelly on a plate, jelly on a plate, wimble wobble, wimble wobble, jelly on a plate
For those of you who thought this weeks post was going to be about eating jelly you are slightly wrong but for those who thought it is about making jelly you are slightly right. This week has involved co-sedimentations. And co-sedimentations require GELS, the scientific equivalent to jelly, at least according to me.
Co-sedimentation is done to see if the components used in the ATPase assays actually bind. If they don't, well then all that time spent pipetting was a bit of a waste. To do this you mix components used in our ATPase assays, in my case this was Actin, tropomyiosn and troponin. These are all components of the thin filament in muscle and they are responsible for regulation of muscle activity. However, to be able to regulate muscle activity they need to be bound to each other. Samples were prepared containing all three components or containing just one component. After a times incubation the samples were run on a gel. Well first the gel had to be made, so to my headline. See making a gel can be slightly compared to making jelly. When you make a gel you mix different solutions, one making it solidify, just like in jelly, hence my headline. But instead of eating our scientific jelly we put our samples on it and ran electricity through it, which in my view is a lot more fun. And what made it even better is that the results showed there was binding, so a big yay my thumb has not been sacrificed in vain
Until next week
Co-sedimentation is done to see if the components used in the ATPase assays actually bind. If they don't, well then all that time spent pipetting was a bit of a waste. To do this you mix components used in our ATPase assays, in my case this was Actin, tropomyiosn and troponin. These are all components of the thin filament in muscle and they are responsible for regulation of muscle activity. However, to be able to regulate muscle activity they need to be bound to each other. Samples were prepared containing all three components or containing just one component. After a times incubation the samples were run on a gel. Well first the gel had to be made, so to my headline. See making a gel can be slightly compared to making jelly. When you make a gel you mix different solutions, one making it solidify, just like in jelly, hence my headline. But instead of eating our scientific jelly we put our samples on it and ran electricity through it, which in my view is a lot more fun. And what made it even better is that the results showed there was binding, so a big yay my thumb has not been sacrificed in vain
Until next week
Sunday 25 July 2010
and we wait
One thing that has been said about science is that when you need time you have non and when you don't you have a lot.
This is exactly what I have been experiencing in my third week here in the lab.
The beginning of the week was definitely a time lacking period. More ATPase assays were on the agenda and if you are slightly absent minded there just isn't enough time to keep those reactions in check. A few mistakes were made and some funny results indeed came out in the end. However, with some proper sleep concentration was returned and what seemed like ok results were obtained, until I ran out of protein that is.
The lack of proteins signified the other part of my week, the one with way to much time. See proteins are extraordinary things but they are also annoying things when they take for ever to be purified. So when you have read all journals imaginary there is nothing left to do than sit and stare at your timer. But all of this is part of being a scientist, well this is my hope.
Until next week, where I hope time will be more evened out
This is exactly what I have been experiencing in my third week here in the lab.
The beginning of the week was definitely a time lacking period. More ATPase assays were on the agenda and if you are slightly absent minded there just isn't enough time to keep those reactions in check. A few mistakes were made and some funny results indeed came out in the end. However, with some proper sleep concentration was returned and what seemed like ok results were obtained, until I ran out of protein that is.
The lack of proteins signified the other part of my week, the one with way to much time. See proteins are extraordinary things but they are also annoying things when they take for ever to be purified. So when you have read all journals imaginary there is nothing left to do than sit and stare at your timer. But all of this is part of being a scientist, well this is my hope.
Until next week, where I hope time will be more evened out
Sunday 18 July 2010
Queen of pipetting
Time really flies in my dear lab, yet another week has passed.
This week all that actin that was purified last week came to good use. The actin was used in several ATPase assays together with myosin, tropomyosin and troponin. Simply explained the ATPase assays are a way of checking the activity of a muscle. The aim is to repeat the assays but with combinations of mutated tropomyosin to see if there is a difference in activity. This difference hopefully explaining different muscle myophaties which is the whole aim with my project.
The assays run this week were mostly practice. In theory it is a simple experiment but in practice you need to be very detailed and good at PIPETTING. If there is one thing I have done this week it is just that, pipetting. I calculated that in one day I pipetted 140 times. Now that gives some thumb cramp. But this weekend my thumb has been on ice so I'm all ready to go tomorrow.
This week all that actin that was purified last week came to good use. The actin was used in several ATPase assays together with myosin, tropomyosin and troponin. Simply explained the ATPase assays are a way of checking the activity of a muscle. The aim is to repeat the assays but with combinations of mutated tropomyosin to see if there is a difference in activity. This difference hopefully explaining different muscle myophaties which is the whole aim with my project.
The assays run this week were mostly practice. In theory it is a simple experiment but in practice you need to be very detailed and good at PIPETTING. If there is one thing I have done this week it is just that, pipetting. I calculated that in one day I pipetted 140 times. Now that gives some thumb cramp. But this weekend my thumb has been on ice so I'm all ready to go tomorrow.
Monday 12 July 2010
one week later
So I am one week into my summer research project and I wish I could say I am now a lab genius but sadly I'm not. However, I have definitely learnt a lot in a short time. In only one week I have seen the purification of actin from beginning to end. And when I mean beginning I mean beginning. It is easy to neglect where protein preps used for experiments come from when doing lab work. Let just put it like this all lab work is not glorious. Hence, I will always appreciate a good actin prep in the future.
The actin which has been purified is going to be used in my future experiments so it is important that it has gone well. Something that lab work doesn't always do. Hence for this week I am keeping my fingers crossed that, even though not a lab genius, my skills have been good enough for the actin to have survived. But if not, at least I now know how to start from the beginning. And we all know what they say practise makes perfect.
The actin which has been purified is going to be used in my future experiments so it is important that it has gone well. Something that lab work doesn't always do. Hence for this week I am keeping my fingers crossed that, even though not a lab genius, my skills have been good enough for the actin to have survived. But if not, at least I now know how to start from the beginning. And we all know what they say practise makes perfect.
Tuesday 6 July 2010
First day!
So today was the start of what will be my 7 weeks in a lab here at Leicester.
Can't say I did much apart from familiarise myself with the place so I don't get lost in the future. But that is always something. Now for some reading and fingers crossed there will actually be lab work tomorrow.
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