Flowchart Southern Blot Labs (2 Days)

Flowchart-Southern blot labs (2 days)

Monday

Traditional blot (gel) / Colony lift
·  Restriction digests (TAs prepared)
·  5 gels (TAs prepared)
·  Run digests on gel at 50mV (1 group per gel), 2 h
·  Make probe (while the gel runs), let stand over night (set up 20 minutes)
·  Prepare gel for blot (1.5 h)
(cut paper towels etc, during the wait) / ·  Colony lift and transfer of DNA onto membrane (30 minutes), auto cross-link (store membrane until Tuesday)
·  Set up over-night blot (10 minutes)

Tuesday afternoon (11am-2pm)

Traditional blot (gel) / Colony lift
·  Dissemble over-night blot and cross link membrane
·  Pre-hybridize both membranes (30 minutes- 1h), and transfer to hybridization solution, leave over night

Wednesday

Traditional blot (gel) / Colony lift
·  Stringency washes (45 minutes)
·  Blocking and detection (2-2.5 h)