Method for developing bidens bipinnata plant SSR (Simple Sequence Repeats) primer based on transcriptome sequencing
A technology of transcriptome sequencing and ghost needle grass, applied in the field of molecular biology, can solve the problem of limited number of SSR primers
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[0089] The SSR primer pair developed based on the transcriptome sequencing of P. figure 1 shown), SSR primer development, DNA extraction, SSR primer screening and variety identification were obtained.
[0090] In the present invention, using the EST sequence obtained by transcriptome sequencing to develop the SSR primer pair for P. sp.
[0091] 1. Transcriptome sequencing of P.
[0092] After extracting the total RNA from the leaves of P. genus plants by the Trizol method, enrich the eukaryotic mRNA with magnetic beads containing Oligo(dT); add fragmentation buffer to fragment the mRNA into short fragments, use the mRNA as a template, and use six bases Random primers (randomhexamers) synthesize the first cDNA strand, then add buffer, dNTPs, RNaseH and DNApolymeraseI to synthesize the second cDNA strand, after purification by QiaQuickPCR kit and elution with EB buffer, do end repair, add poly( A) Connect the sequencing adapter, then use agarose gel electrophoresis to select t...
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