Pooling CASE Block 11 cDNA Libraries for Sequencing

Pooling CASE Block 11 larval oyster cDNA from mRNA Libraries of T24 samples (made in November) for sequencing with Novogene company.

Quants of libraries were calculated from the TapeStation traces by correcting the peak to be between 250 and 600 base pairs and using that concentration (ng/µl)

There are 16 libraries made, and we are pooling into 8 based on index combinations. Index 1 and 2 are paired in individual samples to eliminate the possibility of index hopping. There are two samples per every paired index, each with a different adapter barcode, and we are pooling by the same index pairs.

Sample	Adapter	Index 1	Index 2
B11T24J01	NCO	501	701
B11T24J02	NCO	502	702
B11T24J03	NCO	503	703
B11T24J04	NCO	504	704
B11T24J05	NCO	505	705
B11T24J06	NCO	506	706
B11T24J07	SAII	501	701
B11T24J08	NCO	508	708
B11T24J09	NCO	507	707
B11T24J10	SAII	502	702
B11T24J11	SAII	503	703
B11T24J12	SAII	504	704
B11T24J13	SAII	505	705
B11T24J14	SAII	506	706
B11T24J15	SAII	507	707
B11T24J18	SAII	508	708

Each pool should contain 200ng of library from each sample, and enough 10mM Tris-HCl to add up to 10µl in a screw cap 2mL tube. Tubes were vortexed, spun down, parafilmed on the lid, and shipped with dry ice.

Pool Name	Samples	200ng Vol	Total Pool Vol	Tris-HCl Vol to 10µl	Indexes
CASEB1 pool1	B11T24J01	4.11µl	5.76µl	4.24µl	501 701
	B11T24J07	1.65µl			501 701
CASEB1 pool2	B11T24J02	2.11µl	3.83µl	6.17µl	502 702
	B11T24J10	1.72µl			502 702
CASEB1 pool3	B11T24J03	2.01µl	3.57µl	6.43µl	503 703
	B11T24J11	1.56µl			503 703
CASEB1 pool4	B11T24J04	2.26µl	4.62µl	5.38µl	504 704
	B11T24J12	2.63µl			504 704
CASEB1 pool5	B11T24J05	2.60µl	4.74µl	5.26µl	505 705
	B11T24J13	2.14µl			505 705
CASEB1 pool6	B11T24J06	2.37µl	5.18µl	4.82µl	506 706
	B11T24J14	2.81µl			506 706
CASEB1 pool7	B11T24J09	2.27µl	5.08µl	4.92µl	507 707
	B11T24J15	2.81µl			507 707
CASEB1 pool8	B11T24J08	3.93µl	5.96µl	4.04µl	508 708
	B11T24J18	2.03µl			508 708

Written on March 25, 2019