Construction and screening as well as applications for siRNAs expression carrier of stomach cancer target STAT3 gene
A technology for expressing vectors and genes, applied in the field of screening, gene therapy drugs, and construction of siRNAs expression vectors, can solve the unsatisfactory survival rate of gastric cancer patients and other problems
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[0082] The present invention will be further described below in conjunction with example, but not limited thereto. All reagents and raw materials of the present invention can be purchased through the market.
[0083] 1. Main Reagents
[0084] 1. The vector pGPU6 / GFP / Neo siRNA Expression Vector was purchased from GenePharma (Catalog no: E-07)
[0085] 2. The engineering bacteria JM109 was purchased from Promega (Catalog no: L1001)
[0086] 3. T4 ligase was purchased from NEW ENGLAND (Catalog no: M0202); plasmid extraction and purification kit (QIAprep Spin Miniprep Kit) was purchased from QIAGEN (Cat No: 27104)
[0087] 4. STAT3 and β-actin-specific PCR primer synthesis, STAT3-specific siRNA oligonucleotide chain synthesis and DNA sequence determination (Nanjing GenScript Biotechnology Co., Ltd.)
[0088] 5. Transfection reagent Lipofectamine 2000 (Cat. No: 11668-027, Invitrogen)
[0089] 6. TRzol Reagent (Cat. No. 152476-026, Invitrogen) RT-PCR Kit RNA PCR Kit (AMV) Ver.3....
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