Fluorescent quantitative internal reference genes and their special primers and applications under waterlogging stress in Changshan, Haizhou
A fluorescence quantitative, internal reference gene technology, applied in the field of plant molecular biology, to achieve the effect of improving detection efficiency, strong primer specificity, and improving reliability
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[0030] 1. Plant tissue total RNA extraction and cDNA synthesis
[0031] Extract total RNA with EASYspin Plus Plant RNA Extraction Kit (Adley Biotechnology Co., Ltd.), use a spectrophotometer (METTLER TOLEDO, Switzerland) to detect RNA concentration and quality, and utilize 1.5% agarose gel electrophoresis to detect the total complete RNA. sex. Using Beijing Quanshijin Reverse Transcription Kit (Quanshijin Biotechnology Co., Ltd.), RNA was purified and the first strand of cDNA was synthesized according to the instructions.
[0032] 2. Selection of internal reference genes and design of primers
[0033] Query the main internal reference genes in other plants according to the literature and finally design 17 internal reference genes required by this embodiment, which are Actin (SEQ ID NO.1), PP2A (SEQ ID NO.2), RPL (SEQ ID NO. .3), PK (SEQ ID NO.4), 18S (SEQ ID NO.5), RAN (SEQ ID NO.6), APT (SEQ ID NO.7), SAND (SEQ ID NO.8), PROF (SEQ ID NO. NO.9), MDH (SEQ ID NO.10), EF-1A (S...
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