RNA-Seq of mus musculus:glomerulus
The sequencing library was prepared by truseq nano DNA LT library prep kit of Illumina company.1) First, repair the sequence end of the amplified product, remove the prominent base at the 5 'end of the DNA sequence through the end repair mix 2 in the kit, and add a phosphate group to supplement the missing base at the 3' end;2) Add a base at the 3 'end of the DNA sequence to prevent the self connection of DNA fragments, and ensure that the target sequence can be connected with the sequencing connector (there is a prominent t base at the 3' end of the sequencing connector);3) Add a sequencing connector containing library specific tag (i.e. index sequence) at the 5 'end of the sequence, so that the DNA molecule can be fixed on the flow cell;4) Beckman ampure XP beans was used to remove the self connected fragment of the linker through magnetic bead screening, and the library system after adding the linker was purified;5) PCR amplification was performed on the DNA fragment connected to the connector, so as to enrich the sequencing library template, and the enrichment product of the library was purified again by Beckman ampure XP beans;6) through 2% agarose gel electrophoresis, the final fragment of the library was selected and purified.
A1