Difference between revisions of "20.109(F16): TA notes for M1"
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**10 mL 1x PBS | **10 mL 1x PBS | ||
**2 eppendorf tubes of 0.3% LMP agarose (in eppendorf water bath) | **2 eppendorf tubes of 0.3% LMP agarose (in eppendorf water bath) | ||
| + | **large scheme of the plate so they can keep track of how to load chips | ||
*and separately, in the TC hood, | *and separately, in the TC hood, | ||
**10 mL 1x PBS | **10 mL 1x PBS | ||
| Line 18: | Line 19: | ||
**2 eppendorf tubes of 0.3% LMP agarose (in water bath '''at 43 °C''') | **2 eppendorf tubes of 0.3% LMP agarose (in water bath '''at 43 °C''') | ||
**2x 15 mL of TK6 cells at 500,000 cells/mL | **2x 15 mL of TK6 cells at 500,000 cells/mL | ||
| − | |||
Set up hoods in TC room with, per group: | Set up hoods in TC room with, per group: | ||
*4 binder clips | *4 binder clips | ||
| Line 30: | Line 30: | ||
*team sticker to identify glass slides | *team sticker to identify glass slides | ||
*printout of today's protocol | *printout of today's protocol | ||
| + | *large scheme of the plate to hang in TC so they can keep track of how to load chips | ||
===M1D3=== | ===M1D3=== | ||
Revision as of 18:47, 16 September 2016
M1D1
Per group, aliquot
- 50 mL 1x PBS
- in TC room, 15 mL of “TK6 media”
M1D2
Per group, aliquot
- in main lab
- 10 mL 1x PBS
- 2 eppendorf tubes of 0.3% LMP agarose (in eppendorf water bath)
- large scheme of the plate so they can keep track of how to load chips
- and separately, in the TC hood,
- 10 mL 1x PBS
- 25 mL TK6 media
- 2 eppendorf tubes of 0.3% LMP agarose (in water bath at 43 °C)
- 2x 15 mL of TK6 cells at 500,000 cells/mL
Set up hoods in TC room with, per group:
- 4 binder clips
- 1 glass slide
- 1 bottomless 96-well plate
- 1 razor blade
- Pasteur pipets
- sharp jar
- pipet aid
- pen
- team sticker to identify glass slides
- printout of today's protocol
- large scheme of the plate to hang in TC so they can keep track of how to load chips
M1D3
Per group, aliquot:
- 1% LMP agarose
- H2O2 and MMS
- Alkyline lysis solution
- Alkyline electrophoresis buffer
- Neutralization buffer
- SYBR Gold buffer
M1D4
Per group, aliquot:
- 15 mL 1x PBS
- H2O2
- Alkyline lysis solution
- Warm M059J/K culture medium
M1D5
Per group, aliquot in TC room:
- 5 mL of 0.1% gelatin
- 10 mL of PBS
- 3 mL of 1X trypsin
- 15 mL of M059J/K media
- M059J/K in T25 flasks
M1D6
Per group, aliquot in TC room:
- 10 mL of 1x TBS
- 500 uL of 10% BSA, frozen stock in -20C
- Triton X-100 at front bench
- Primary antibodies
M1D7
Per group, aliquot
- 5 mL of 1x TBS
