title: ns192
design: Animals were grown on E. coli strain OP50 until bacteria were depleted, harvested in M9, and resuspended in 1 M pH 7.5 TEN. SDS and proteinase K were added to a final concentration of 0.5% and 0.1 mg/ml respectively and 1 l of -mercaptoethanol was also added. The reaction was incubated overnight in a shaking thermocycler at 56C. Phenol/chloroform was added and phase-separated by spinning in a phase-lock tube. The aqueous phase was transferred to 200 proof EtOH. The resulting DNA clot was washed in 70% EtOH, dried, and resuspended in TEN. After rehydration, 0.3 l of 100 mg/ml RNAse A was added and incubated at 37C for 2 hours. Phenol/chloroform extraction was performed again and DNA was rehydrated with EB buffer. The sample was then run on a nanodrop machine and on a 1% gel to confirm that there was no RNA contamination or DNA degradation, respectively. NextSeq High Output 1 x 75 sequencing was performed with Standard Illumina Sequencing primers for gDNA-seq application.