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目的探讨普通甲醛对人体组织DNA的影响及提高STR检出率的手段。方法取少量石蜡包埋组织,65℃水浴脱蜡,Chelex-100法提取DNA,PCR扩增,循环次数分别为28次和28+6次,扩增产物经310型测序检测。结果普通甲醛固定5d以内的组织基本上可检出Amel及15个STR基因座,固定时间延长,检出率下降;PCR循环次数增加,灵敏度增加,但有错误分型的情况。结论普通甲醛固定时间长短直接影响PCR-STR的检验,减少模板量有利于PCR反应成功,PCR次数为28+6时,灵敏度增加,但应谨慎判读结果,以免错误分型。
Objective To investigate the effect of common formaldehyde on the DNA of human tissues and the means of increasing the detection rate of STR. Methods A small amount of paraffin embedded tissue was obtained and dewaxed in a water bath at 65 ° C. DNA was extracted by Chelex-100 method. The PCR amplification was carried out with 28 cycles and 28 + 6 cycles, respectively. Results The results showed that Amel and 15 STR loci were detected basically within 5 days of normal formaldehyde fixation. The fixation time was prolonged and the detection rate was decreased. The number of PCR cycles increased and the sensitivity increased, but there was an error in typing. Conclusion The regular formaldehyde fixation time directly affects the test of PCR-STR. Reducing the amount of template is favorable for the success of PCR reaction. When the PCR number is 28 + 6, the sensitivity increases, but the result should be carefully interpreted to avoid wrong typing.