Reagent kit and method for detecting human alpha defensin 1/3 gene copy number
A technology of gene copy number and defensin, applied in the field of biological diagnosis, can solve the problems of high cost and long time
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[0030] A kit for detecting the copy number of DEFA1 / DEFA3, the main reagents include: specific fluorescent quantitative PCR primers (4μM / μl) for detecting DEFA1 / DEFA3 sites; specific DEFA1 / DEFA3 probes; internal reference gene ALB specific fluorescent quantitative PCR primer (4μM / μl); specific ALB probe; check sample DNA; positive control DNA; fluorescent quantitative PCR reaction solution.
[0031]According to the corresponding sequences of the internal reference gene ALB and the target gene DEFA1 / DEFA3 in GenBank (number: ALB is NM000477.5, DEFA1 is NM004084.3, DEFA3 is NM005217.3), select a conserved fragment with a length of 50-150bp, and use Primer Express 2.0 The software designs specific probes and primers. The specific probe length is between 16-25 bases, the GC content is 30-80%, the C content is higher than the G content, and the annealing temperature (Tm value) is 65-70°C, avoiding stable dimers and hairpins structure. Design 3-4 pairs of suitable intron-specific ...
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