Multiple PCR reagent kit detecting breast cancer susceptibility gene mutation and preparation method thereof
A kit and breast cancer technology, which is applied in the field of medical molecular biology, can solve the problem of high prevalence of breast cancer and achieve the effects of avoiding complicated procedures, strong sensitivity and high accuracy
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[0028] Embodiment 1, preparation and assembly of the multiplex PCR kit of the present invention
[0029] 1. Design and preparation of primer pair 1 and primer pair 2
[0030] Primer pair 1: BRCA1 F: 5-TAC CCAGTT GGT TGATTT CC-3 (SEQ ID NO: 1)
[0031] R: 5-CTC ACACAG GGGATC AGC ATT C-3 (SEQ ID NO: 2)
[0032] Primer pair 2: BRCA2 F: 5-GGG AAG CTT CAT AAG TCA GTC-3 (SEQ ID NO: 3)
[0033] R: 5-TTT GTAATG AAG CAT CTG ATA CC-3 (SEQ ID NO: 4)
[0034] It was synthesized by an automatic DNA synthesizer and diluted to 50 pmol / μl.
[0035] 2. Assembly of kit
[0036] 10×PCR reaction buffer 0.002-10ml, dNTPs 0.025-10ml, 25mmol / L MgCl 2 0.002ml-10ml; heat-resistant DNA polymerase 0.3μL-1500μL; primer pair 0.5μL-1500μL; primer pair 20.5μL-1500μL, double distilled water 20μL-100mL; BRCA1 exon 11 positive control 15μl-5mL, BRCA2 first Exon 11 positive control 15μl-5mL, BRCA1 exon 11 negative control 15μl-5mL, BRCA2 exon 11 negative control 15μl-5mL.
[0037] Among t...
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