Thursday, February 12, 2015

bio125, Thu, PCR reaction, past exams, restriction enzyme lab report

8-8:45am Go over PCR protocols, using R code for dilution. Make master screen cast.
Group 2 worked out the master mix on the board.

Each rxn: 2 uL of primer mixture and 23uL water for each reaction + 1 bead
                 2 uL 5n/uL template

8:45-9:30am. students diluted their plasmid stock solution to 5ng/uL as PCR templates.

by 9:30 Setup PCR. (Warning: Glove and PCR bead can attract with electrostatic pull)
First group finished setting up PCR, but some groups still work on the template dilutions.

I asked the leading group to follow youtube video and work on primer analysis.

Screen cast:
PCR primers on pMSH2, followed by restriction enzymes
Problem: some students choose 'circuar' for PCR framgment.
ApE PCR primer analysis and followed by restriction enzyme, gel simulation

 Go over lab report:  RE lab report will be due, video and R code

Only five students were in by 1:10pm.

1:15pm start class.

2:10pm students started setting up reactions.

by 3pm. video to use provided RE to analyze PCR fragments of WT and msh2 mutants.

Unused and Moved to next week

MCAT PCR questions

PCR_inclass_data analysis

Past exam questions on gel pictures.

todl Design site-directed mutagenesis primers

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