Project: Thermal Transplant Molecular

Goal

Fourth 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 #
18-227	R-11-A	Adult	15	15
18-370	P-15-A	Adult	16	16
18-130	R-5-A	Adult	17	17
L-1263	R-7-A	Larvae	18	18
L-924	P-9-A	Larvae	19	19
L-562	R-5-A	Larvae	20	20
L-1059	R-15-A	Larvae	21	21
18-67	P-14-B	Adult	22	22

Qubit Results

To test DNA quantity: Qubit

I used the DNA broad range assay.

I ran the qubit and tapestation on the following day (20210427). After the bead clean up step I stored the libraries in the fridge at 4C for ~16 hours.

Sample	DNA 1 (ng/ul)	DNA 2 (ng/ul)
Standard 1	232.77
Standard 2	19299.53
15	LOW	LOW
16	21.0	20.2
17	5.94	5.68
18	14.3	14.2
19	LOW	LOW
20	3.36	3.12
21	4.74	4.64
22	19.3	18.6

Tapestation Results

D5000 Tapestation protocol

Full Tapestation Report