In order to determine if ApeKI restriction enzyme will work for O. lurida, did test digestion with 4 individuals from different populations. Used ApeKI from Winston.
Test digestion with ApeKI
- Determine concentration with Qubit (did not update standard from 11/5))
- OR1_2: 4.78 ng/ul
- W13_1: 6.82
- BC4_17: 9.62
- CA1_17: 96.4
- Dry down in CentriVac at 60, ul for 200 ng:
- B: 20.79
- C: 2.07
- O: 41.84
- W: 29.32
- ApeKI digestion at 75deg
- 17 ul dH2O, 2 ul NEB3, 1ul ApeKI
- 2:10p until 4:15pm
Sequencing Reaction of Sap/Exo
- 86 reactions
- well 16 from SapExo was empty (did not put lid on tight enough?)
- BC3_11
- well 16 from SapExo was empty (did not put lid on tight enough?)
- Used cyc-seq program from Barber
Getting My Genes Wet 