20.109(S08): TA notes for module 2

General notes

Key preparation:

• Lots of wild-type inverse pericam plasmid (in the pRSET vector) must be available.

Daily Notes

Day 1

Materials required:

1. None: all work today is computer work.

Day of Lab:

• No quiz.
• Primers for mutagenesis must be ordered right away!

Day 2

Materials required:

1. Quick-Change SDM kit. ?1/2? reactions per student, plus 1 control reaction, plus spare reagents (ideally).
   • cat # 200519
   • e.g., for 12 pairs and 1 mutant per pair, two 10 rxn kits would suffice; for 12 pairs and 2 mutants per pair, 1 30 rxn. kit should suffice.
2. sterile DI water
3. LB+Amp plates (at least 30)
4. autoclaved glass tubes (at least 50)
5. LB broth, ampicillin
6. competent XL1-Blue cells

Day of Lab:

• Quiz (prepared by TA).
• Guide journal article discussion (assign figures at beginning of class).
• At end of day: transform mutant DNA, after digesting parental plasmid, into competent XL1-Blue cells.