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

RRBS

• Followed digest and size selection plan exactly as outlined here
  • to get 1ug of DNA in 20uL for each sample, I followed this doc containing the initial sample concentrations
• 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