Tues. Jul. 9, 2019 Salmon RRBS + Zymo Pico Methyl prep

RRBS

  • Followed digest and size selection plan exactly as outlined here
  • DNA yield after digestion and size selection ranged from 20-110ng, which is on par with what I got in the June 28 test run
    • concentration and yields can be found here
    • Digested size selceted samples (tubes labelled “sz gDNA 1-20”) are stored in 209 -20C bottom drawer in box labeled “gDNA Salmon Skin sea lice infection (from Christian) S.Trigg”

Zymo Pico Methyl kit prep

  • picked up primers today from Biochem stores
  • reconstituted in EB (Qiagen) @ 100uM
  • added them to the primer stocks inventory
  • prepared 50uL of 10uM Index primer mixes following Karolyn’s instructions:
    • 50uL Index Primer 10uM (section 5) =
      • 5uL 100uM Ill_TrSq_P5_Univ
      • 5uL 100uM Ill_TrSq_P7_# (where # = index 1-22)
      • 40uL EB
    • We now have Index Primer mixes 1-22 with #2,4,5,6,7,and 12 coming from the kit and all others coming from IDT/I made.

Plan for tomorrow

  • Set up programs on PCR machine
  • Aliquot out up to 50ng Salmon DNA (see volumes here)
  • Aliquot out 50ng of sea lice DNA
  • start library prep
Written on July 9, 2019