DNA Isolation - Claire’s C.gigas Female Gonad for Illumina Bisulfite Sequencing

Due to poor “tag counts” from the initial sequencing (DATE) and the re-sequencing (20131127) of this sample, the HTGU facility has concluded that the library is probably at fault. They will make a new library and do a quality control run on the new library. However, they have insufficient gDNA left to make a new library.

Isolated gDNA from Claire’s sample following the DNAzol protocol.

Transferred ~300uL of female C.gigas gonad from the source tube (ethanol-preserved) to a clean tube. Pelleted gonadal material by spinning 10,000g, 30seconds, @ RT. Decanted residual ethanol. Resuspended tissue in 500uL of DNAzol + 100ug of Proteinase K (Fermentas; 18.5mg/mL). Incubated on a rotator for ~6hrs. Proceeded according to DNAzol protocol. Resuspended final pellet in 100uL of Elution Buffer (Qiagen; EB). After resuspension, pelleted remaining debris 16,000g, 30seconds, @ RT. Transferred supernatant to clean tube and quantified on NanoDrop 1000.

CgF - 403.2ng/uL

Will bring tube to sequencing facility tomorrow morning.