Difference between revisions of "20.109(S08): TA notes for module 2"
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*No quiz. | *No quiz. | ||
*Primers for mutagenesis must be ordered right away! | *Primers for mutagenesis must be ordered right away! | ||
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| + | ===[[20.109(S08):Site-directed_mutagenesis (Day2) | Day 2]]=== | ||
| + | |||
| + | '''Materials required:''' | ||
| + | |||
| + | #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. | ||
| + | #sterile DI water | ||
| + | #LB+Amp plates (at least 30) | ||
| + | #autoclaved glass tubes (at least 50) | ||
| + | #LB broth, ampicillin | ||
| + | #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. | ||
Revision as of 19:45, 3 January 2008
Contents
General notes
Key preparation:
- Lots of wild-type inverse pericam plasmid (in the pRSET vector) must be available.
Daily Notes
Day 1
Materials required:
- None: all work today is computer work.
Day of Lab:
- No quiz.
- Primers for mutagenesis must be ordered right away!
Day 2
Materials required:
- 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.
- sterile DI water
- LB+Amp plates (at least 30)
- autoclaved glass tubes (at least 50)
- LB broth, ampicillin
- 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.
