Bio125, Feb 11, Tue, 2014, ApE, identify mutations, gel electrophoresis

I manually generated mutations and give students for analysis.  I made two mistakes and were found out by studetns.

I posted a youtube tutorial and asked student finish the ApE assignment in class. This seems to push student to focus on the work. At 1:30pm, the college was closed and some student left. Others stay and kept working on the ApE exercieses.

Gel were poured by lab instructor. Student loaded their samples, and gels were run. I found that ApaI and Xab I gave two similar-sized bands for pMSH2 and one sized-band for pRS413. When the plasmid are overloaded, the one 4.5kb smear band is similar to the 4.4 and 4.9 kb bands. We decided to run the gel longer to get better resolution.  (Next time we should pick different RE, or we can ask student to pick better RE. 0.8% gel might be used). 



Because of the HA tag, locations can be different between MSH2 and  HA-MSH2, (info based on D. Wang). 

1.     G704S: AG41_G693S

2.     R542P: AG29_R542P

3.     H669Y: AG33_H658Y

4.     G704D: AG91_G693D

5.     M718I: AG371_M707I

6.     H669R: AG372_H658R

7.     A618V: AG421_A618V

8.     D621N: AG422_D621N