7:30 am – 10:00 pm
(9:30 am – 11:30 am volleyball)
RT reactions
Probes in pipeline
- L2 F01 – F07
- L3 E06 – E09
- making with both P2 and P3
Per RT
- 15 uL RNA
- 4 uL primer
- 6 uL RT buffer
- 2 uL dNTPs
- .5 uL RNAsin
- 2 uL ddH2O
- 0.5 uL Maxima
- Total volume = 30 uL reaction
12 reactions master
- 48 uL primer
- 72 uL RT buffer
- 24 dNTPs
- 6 uL RNasin
- 6 uL Maxima
- 12 uL ddH2O
- now OUT of Maxima
- now OUT of RNasin
digestion and cleanup
- added 2x volume of EDTA + NaOH (equal volume would have been sufficient)
- volume for cleanup now greater than 1 column
- accidently starting adding on P2 probes E6 through E9 in backwards order (should have kept this in ordinal order at the T7 stage).
- Lib
Test gels
- all my precast gels have been used
- started casting new gels
- acrylamide won’t polymerize today
- added more TMED and APS, still no polymerization
Cell staining
- new stains:
- F06-p2-A647 + F07-p3-A750
- F07-p2-A647 + F06-p3-A50