Quantitative detection method of HPP1 gene methylation
A technology of methylation quantification and detection method, which is applied in the field of HPP1 gene methylation detection, can solve the problems of complexity, large amount of samples, and restrictions on the use of MSP, and achieve high sensitivity, high accuracy, and easy operation Effect
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[0021] Example 1: Quantitative detection of methylation degree of HPP1 gene (GeneID: 23671) in clinical samples of patients
[0022] Sources of main reagents and instruments: QIAGEN EpiTect Bisufite KitTM kit, pMD18-TVector and EX-taq enzymes were purchased from Treasure Bioengineering (Dalian) Co., Ltd., ABI Taqman GeneExpression Master Mix was purchased from Applied Biosystems, Inc., and ABI 7500 Real Time PCR instrument Primers were synthesized by Shanghai Bioengineering Technology Service Co., Ltd. in Applied Biosystems, USA.
[0023] 1. Preparation of standard products using gene amplification and bioengineering methods
[0024] 1. Genomic DNA Extraction
[0025] DNA was extracted by conventional phenol / chloroform method.
[0026] (1) Put 50mg of tissue into a 5ml centrifuge tube, add 700ul of lysis buffer to make a homogenate.
[0027] (2) Add proteinase K to a final concentration of 100ug / ml, and keep at 55°C overnight until the solution becomes clear.
[0028] (3) ...
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