Gene immunity preparation method for human I-type deoxythymidine kinase monoclonal antibody
A glycoside kinase monoclonal antibody technology, applied in botany equipment and methods, biochemical equipment and methods, genetic engineering, etc., can solve the problems of miniscule, long cycle, and difficult problems, and achieve easy unification, standardization, and high specificity The effect of sex and easy operation
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[0026] Examples:
[0027] ①Cloning and sequence analysis of hTK1 gene:
[0028] 1) Reverse transcription PCR amplification of TK1 cDNA: Trizol reagent kit, which can extract total ribonucleic acid (RNA) from animal cells and tissues, bacteria, fungi, etc., was used to extract RNA from 5 million Hela cells and treated with methotrexate 48 After purification, use a high-efficiency alkylating agent diethyl pyrocarbonate (DEPC) to dissolve the precipitate in water to obtain 30μl. Because DEPC can destroy the activity of ribonuclease (RNase), it can remove RNase contamination.
[0029] When performing reverse transcription, add 5 times buffer: 4μl, primer Oligo-dT (0.5μg / 1μl)
[0030] 1μl, 10mMol / L deoxynucleoside triphosphate (dNTP): 2μl, RNA: 10μl, deionized water: 2μl, mix thoroughly, and place in a constant temperature water bath at 70℃ for 10 minutes; then, place on ice 2 minutes; add 1μl of reverse transcriptase (50U, Roche), mix well, and place in a water bath at 42°C for 90 min...
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