Additional Testing

Welcome back everyone! This week was all about testing, re-testing, and even more testing of the pfu I created. Of course, this testing involved the use of PCR, which can be quite aggravating to set up, but some more definitive results are starting to appear. I have managed to find the proper concentration of my pfu for PCR, so all that remains is finding other possible buffers to make it run more efficiently or adding other proteins to make the pfu more stable.

As for the proper concentration, I have found that .25µL of my stock pfu works very well in 25µL of solution. I have even been able to compare these results with that of taq polymerase (another thermostable DNA polymerase and the one my project was originally going to be about) and they seem about even in the amount of DNA produced through PCR, see my last post for a picture.

I have now been experimenting with different buffers, which did not work at all, but we kind of scrapped the idea of using new buffers as my first one seems to work just fine. So, now we are experimenting with adding a protein to the solution called BSA, Bovine Serum Albumin. What BSA is meant to do is provide stability to the pfu which causes it to function better. Unfortunately, for all the PCR tests I have done with it, I have gotten pretty inconsistent results, whether that's my fault (which it probably is) or some other problem, its hard to say. Sometimes it works far better than without it, other times it makes the PCR not work at all. I will likely test this again next week, to find out for certain if it really does do anything.

Since I do not have any very interesting pictures other than a ton of pictures of all the different PCRs I have done, I thought I might show you what the bench I work at looks like.

Ooooooooh Science!

 And in case you were curious, this is what it looks like to run a gel electrophoresis as that is how I know how well my PCR turned out.

More Science!

I am nearing the end of this project and I thank all of you for reading it up until now, if you haven't well better late than never! See you all next week!