20.109(F16): TA notes for M1

For Hsinhwa

Module 1 Measuring Genomic Instability Number of groups: 9, so prepare 11 aliquots

Week 1: September 12, 2016 Day 1 (T/W) Per group: • Aliquot 50 mL 1x PBS per group (label: PBS) • In TC room, aliquot 15 mL of “TK6 media” (label: TK6 media)

Day 2 (R/F) Per group: • Aliquot 10 mL 1x PBS (label: PBS) • And separately, in the TC hood, aliquot 10 mL 1x PBS (label: PBS) Leslie & Maxine will prepare: • Prepare 4 x 2 mL tubes of 0.3% LMP agarose, per group

Week 2: September 19, 2016 Day 3 (T/W) Leslie & Maxine will prepare: • 1% LMP agarose • H2O2 and MMS • Alkyline lysis solution • Alkyline electrophoresis buffer • Neutralization buffer • SYBR Gold buffer

Week 3: September 26, 2016 Day 4 (T/W) Per group: • Aliquot 15 mL 1x PBS (label: PBS) Leslie & Maxine will prepare: • H2O2 • Alkyline lysis solution • Warm culture medium

Day 5 (R/F) Per group: In TC room for all aliquots, leave at 4C: • Aliquot 5 mL of 0.1% gelatin (label: Gelatin) • Aliquot 10 mL of PBS (label: PBS) • Aliquot 3 mL of 1X trypsin (label: Trypsin) • Aliquot 15 mL of M059J/K media (label: Media) Leslie & Maxine will prepare: • M059J/K in T25 flasks

Week 4: October 3, 2016 Day 6 (T/W) Per group: • Aliquot 10 mL of 1x TBS (label: TBS) • Aliquot 500 uL of 10% BSA, frozen stock in -20C (label: 10% BSA) Leslie & Maxine will prepare: • 10% BSA and freeze at -20C • Triton X-100 at front bench • Primary antibodies

Day 7 (R/F) Per group: • Aliquot 5 mL of 1x TBS (label: TBS)