20210504 WGBS PicoMethyl-seq library prep for Thermal Transplant Trial 6

Project: Thermal Transplant Molecular

Goal

Sixth trial of WGBS library prep using the Zymo Pico Methyl-seq library prep kit and this protocol.

Sample dilution and mix calculations can be found on this google sheet

Protocol

I am using a modified version of this protocol.

For all samples, I used the following modifications:

  • Use the combined i5 and i7 index primers (an input of 1uL)
  • Reduce the Library Amp Master Mix (2x) to 13uL (was originally 14uL)

Samples and Indices

Sample Coral ID Life Stage i5 Index # i7 Index #
18-239 R-8-B Adult 33 33
18-190 P-14-A Adult 34 34
18-311 P-10-B Adult 35 35
18-202 P-4-A Adult 36 36
18-142 R-8-A Adult 37 37
18-55 R-7-A Adult 38 38
18-178 R-5-B Adult 39 39
18-394 P-9-B Adult 40 40

Qubit Results

To test DNA quantity: Qubit

I used the DNA broad range assay.

Sample DNA 1 (ng/ul) DNA 2 (ng/ul)
Standard 1 181.18  
Standard 2 20085.23  
33 8.82 8.70
34 23.0 22.8
35 21.0 20.8
36 25.0 25.0
37 20.2 19.6
38 22.6 22.4
39 14.1 14.1
40 17.6 17.6

Tapestation Results

D5000 Tapestation protocol

Full Tapestation Report

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Written on May 4, 2021