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After amplification and purification, VAHTS Universal DNA Library Prep Kit for Illumina V3 was used to construct library. The input-DNA was quantified by Qubit2.0, and the size was measured with Qsep100 DNA Fragment Analyzer. Then end-repair and adenylation were performed. The reaction mixture containing fragmented DNA (50 ng), end repair buffer, end repair enzymes, and nuclease-free water was incubated at 30°C for 30 minutes and inactivated at 65°C for 30 minutes. The finished end prep reaction mixture was added with working adaptor and ligation enzymes and then was incubated at 20°C for 15 minutes. The ligated DNA was purified, and size was selected with AMPure XP beads. The library amplification was followed, and purification was performed with AMPure XP beads. The final library was quantified by Qubit2.0 and the library size was measured with Qsep100 DNA Fragment Analyzer. Library sequencing was performed using the NovaSeq 6000 and S4 Reagent Kit with paired end reads of 150. |
RNA-Seq |
TRANSCRIPTOMIC SINGLE CELL |
RANDOM |
PAIRED
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