Difference between revisions of "20.109(F16): TA notes for M2"
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MAXINE JONAS (Talk | contribs) (→M2D1) |
MAXINE JONAS (Talk | contribs) (→M2D2) |
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Fa16: all done by 4:30pm | Fa16: all done by 4:30pm | ||
'''Should schedule BE Comm Lab workshop on Journal Club on this day.''' | '''Should schedule BE Comm Lab workshop on Journal Club on this day.''' | ||
| + | |||
| + | ===M2D3=== | ||
| + | On ice, | ||
| + | *1.2 mL aliquots of nuclease-free water | ||
| + | *reverse primer at 100 μM | ||
| + | *psgRNA template | ||
| + | *HotStart Master Mix | ||
| + | *-20 °C block for PCR tubes | ||
| + | *strip of PCR tubes with colors (+1 for control) | ||
| + | *SDM control primer mix | ||
| + | *SDM control plasmid | ||
| + | |||
| + | Also on front bench, | ||
| + | *filtered tips | ||
| + | *gRNA forward primers received from IDT | ||
Revision as of 17:45, 20 October 2016
M2D1
Per group, aliquot
- 30 μL nuclease-free water
- 5 μL pdCas9 (ideally at 500 ng/μL)
On front bench, have
- restriction enzymes in -20 °C block
- NEB buffers
- list of 20.109-owned restriction enzymes
Fa16: all done by 5pm
M2D2
Per group,
- pour 1/2 agarose gel (only 5 lanes needed)
- thaw 4 samples of confirmation digests
- aliquot 10 μL of 6X loading buffer
- thaw 20 μL of 1 kb ladder
Call IDT to request expedited FedEx shipping of gRNA oligos.
Fa16: all done by 4:30pm Should schedule BE Comm Lab workshop on Journal Club on this day.
M2D3
On ice,
- 1.2 mL aliquots of nuclease-free water
- reverse primer at 100 μM
- psgRNA template
- HotStart Master Mix
- -20 °C block for PCR tubes
- strip of PCR tubes with colors (+1 for control)
- SDM control primer mix
- SDM control plasmid
Also on front bench,
- filtered tips
- gRNA forward primers received from IDT
