Fluorescent quantitative internal reference genes and their primers and applications in different tissues of Cinnamomum camphorensis
A fluorescence quantitative, internal reference gene technology, applied in the application, genetic engineering, plant gene improvement and other directions, can solve problems such as quantitative research on the relative expression of pest functional genes, and achieve the effect of high amplification efficiency and strong specificity
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[0026] 1. Extraction of total RNA and synthesis of cDNA from different tissues of Cinnamomum chinensis
[0027] Different tissues of 4th instar larvae and adults were collected (the head does not contain antennae, thorax, abdomen, wings, feet, and antennae). Each treatment had three biological replicates, and they were frozen in liquid nitrogen, and the Trizol method was used after treatment. Total RNA was extracted and treated with 5 mL of RNase-free DNase I (Takara Bio Inc., Kusatsu, Shiga Prefecture, Japan) for 15 minutes to remove DNA. The concentration and quality of RNA were detected using a Thermo Scientific NanoDrop 2000 UV-Vis spectrophotometer (Thermo FisherScientific Inc., Waltham, MA, USA). The integrity of total RNA was detected by 1.0% agarose gel electrophoresis. Using PrimeScript TM II first-strand cDNA synthesis kit (TaKaRa, Dalian, China), purify RNA and synthesize the first-strand cDNA according to the instructions. Perform reverse transcription in a 20.0...
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