Project: Thermal Transplant Molecular

Goal

Fifth 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 #
18-406	R-15-A	Adult	23	23
18-32	P-12-B	Adult	24	24
18-262	P-15-B	Adult	25	25
18-322	P-9-A	Adult	26	26
18-79	R-14-A	Adult	27	27
L-661	R-9-A	Larvae	28	28
L-1029	P-4-A	Larvae	29	29
L-862	P-14-A	Larvae	30	30
L-1038	R-8-A	Larvae	31	31
L-1257	P-19-A	Larvae	32	32

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	177.01
Standard 2	19683.34
23	24.0	23.6
24	13.1	12.5
25	15.2	14.8
26	3.68	3.40
27	5.64	5.24
28	3.16	2.56
29	7.84	7.54
30	LOW	LOW
31	14.2	13.9
32	5.78	5.36

Tapestation Results

D5000 Tapestation protocol

Full Tapestation Report