GSM5224486: CCM3_Ammo_30; Homo sapiens; OTHER
Confluent cells from 6 well dishes were scraped from the plate, pelleted at 800 g for 5 min, and genomic DNA was extracted using the Qiagen DNAeasy kit per manufactures protocols, and eluted in 100 μL of low EDTA TE buffer. The transgene region was amplified using primers targeted to the tetracycline promoter (located in the attP landing pad) and the IRES site (located in the attB plasmid), such that only successful integrated transgenes were amplified. PCR was carried out at 100 μL scale using NEB Q5 high fidelity polymerase per manufacturers protocols using 5 μL of genomic DNA, an annealing temperature of 68 °C, and a 3 min extension time to limit bias towards shorter products (ATP5C) for a total of 30 cycles. PCR amplicons were purified using the NEB Monarch PCR clean-up kit per manufactures protocols. Nextera Flex library prepration on PCR amplicons
GEO Accession
GSM5224486