ATAC_IEC_Week_2_3
Nuclei are extracted from samples and tested for quality control (QC). After the QC, the nuclei pellets are resuspended in the Tn5 transposase reaction mix, which includes transposase and equimolar Adapter1 and Adapter 2. The mix of transposition reaction is incubated at 37C for 30 min. PCR is then performed to amplify the library. Libraries were purified with the AMPure beads and library quality was assessed. After the construction of the library, the initial quantification was done with Qubit 2.0, and the library is diluted to 1ng/l. Then the insertion size of the library is detected with NGS3K. After library quality control, sequencing is performed for different libraries according to the concentration and the demand of data amount on Illumina HiSeq/NovaSeq platform. The basic principle of sequencing is sequencing by synthesis. Four kinds of fluorescent labeled dNTP, DNA polymerase and adapter primers are added to the flow cell for amplification. When each sequence cluster extends the complementary chain, each fluorescent labeled dNTP releases the corresponding fluorescence. The sequencer captures the fluorescent signal and converts the light signal into the sequencing peak through computer software to obtain the sequence information of the fragment to be detected.