Project: Thermal Transplant Molecular

Goal

Seventh trial of WGBS library prep using the Zymo Pico Methyl-seq library prep kit and this protocol. I finished the first prep of every sample and re-did one sample that previously failed (i.e. too low to read on the Qubit, L-862).

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 #
18-346	P-6-B	Adult	41	41
L-571	R-11-A	Larvae	42	42
18-250	R-11-B	Adult	43	43
18-418	R-9-A	Adult	44	44
18-454	P-12-A	Adult	45	45
18-44	R-19-B	Adult	46	46
18-466	R-15-B	Adult	47	47
L-862	P-14-A	Larvae	48	48

L-862 was the sample I re-did.

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	184.29
Standard 2	20541.44
41	7.28	6.94
42	10.1	10.2
43	8.22	8.18
44	21.2	21.0
45	5.42	5.38
46	21.6	21.4
47	19.1	19.0
48	24.4	24.2

Tapestation Results

D5000 Tapestation protocol

Full Tapestation Report