Haemaphysalis Iongicornis transcriptome sequencing
After extract total RNA and treated with DNase I, Oligo(dT) are used to isolate mRNA. Mixed with the fragmentation buffer, the mRNA are fragmented. Then cDNA is synthesized using the mRNA fragments as templates. Short fragments are purified and resolved with EB buffer for end reparation and single nucleotide A (adenine) addition. After that, the short fragments are connected with adapters. The desired fragments are selected for the PCR amplification. During the QC steps, Agilent 2100 Bioanaylzer and Qubit 2.0 Fluorometer are used in quantification and qualification of the sample library. Then the library is sequenced using Illumina HiSeq X10.
SUB2801738
Haemaphysalis Iongicornis:XJTU Zhao lab