Please help for SNP detection# Biology - 生物学
L*e
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Using TaqMan SNP genotyping assay to detect a SNP. The problem is that all
samples (blood genomic DNA) always showed the same "heterozygous" patten:
signals for WT and SNP are the same.
Does that mean the probes bind unspecifically? i tried different annealing
tep. at 60, 62 and 64 C degree, looks no much change.
The Tm of the primers: 55 C (forward) and 59 C (reverse)
The Tm of probes: 51 C (WT), 49 C (SNP)
PCR product: 90 bps
The primers and probes are designed by the ABI.
What can i do next? Thanks.
samples (blood genomic DNA) always showed the same "heterozygous" patten:
signals for WT and SNP are the same.
Does that mean the probes bind unspecifically? i tried different annealing
tep. at 60, 62 and 64 C degree, looks no much change.
The Tm of the primers: 55 C (forward) and 59 C (reverse)
The Tm of probes: 51 C (WT), 49 C (SNP)
PCR product: 90 bps
The primers and probes are designed by the ABI.
What can i do next? Thanks.