GSM4258546: 190805-Index26; Mus musculus; OTHER
Genome DNA isolation of transfected N2a cells and FACS collected cells were done by QuickExtract™ DNA Extraction Solution 1.0 (Lucigen) following the manufacturer's instructions. For each targeting genome locus, an amplicon of 100-300 bp were designed. Two-runs of PCR are applied to ligate adaptors and index for following deep sequencing. Firstly, the first PCR amplified sites of interest with primers containing IIlumina adaptors. And exonuclease I was added to the PCR1 products to clear all remaining PCR1 primers. Then a second run of PCR was performed to add unique barcodes and P5/P7. All the PCR reactions were carried out with Kapa HiFi 2 x mastermix (Kapa Biosystems). PCR products were then separate by 2% agarose gel, only products with desired sizes were cut and purified by HiPure Gel Pure DNA mini kit (Magen).