GSM5262227: Ex52_1to1_mouse1_local (gDNA); Mus musculus; OTHER
Tissue was extracted from euthanized mice. Genomic DNA was isolated using the DNeasy kit (Qiagen) according to the manufacturer's protocol. Total RNA was isolated using QIAshredder and RNeasy Plus kits (Qiagen). First-strand cDNA synthesis was performed using 500 ng total RNA per sample using the SuperScript VILO Reverse Transcription Kit (Invitrogen) and second-strand synthesis was performed using Klenow fragment DNA polymerase (NEB). Tagmentation of 200 ng genomic DNA or second-strand products was performed using a 1:40 dilution of assembled Tn5. To enrich the targeted sequence, first round PCR using a genome specific primer was used with a reverse primer specific for the i7 adapter sequence inserted by the transposon. Second round PCR using a barcode primer specific for the custom adapter sequence was used to add 6-nucleotide experimental barcodes and the Illumina i5 adapter was used with the Tn5-Universal reverse primer. Tn5-based tagmentation with target enrichment.
GEO Accession
GSM5262227