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在超薄平板凝胶中分离的聚合酶链反应(PCR)产物的毛细管进样。

Capillary sample introduction of polymerase chain reaction (PCR) products separated in ultrathin slab gels.

作者信息

Bullard K M, Hietpas P B, Ewing A G

机构信息

Department of Chemistry, The Pennsylvania State University, University Park 16802, USA.

出版信息

Electrophoresis. 1998 Jan;19(1):71-5. doi: 10.1002/elps.1150190113.

Abstract

Polymerase chain reaction (PCR) amplified short tandem repeat (STR) samples from the HUMVWF locus have been analyzed using a unique sample introduction and separation technique. A single capillary is used to transfer samples onto an ultrathin slab gel (57 microm thin). This ultrathin nondenaturing polyacrylamide gel is used to separate the amplified fragments, and laser-induced fluorescence with ethidium bromide is used for detection. The feasibility of performing STR analysis using this system has been investigated by examining the reproducibility for repeated samples. Reproducibility is examined by comparing the migration of the 14 and 17 HUMVWF alleles on three consecutive separations on the ultrathin slab gel. Using one locus, separations match in migration time with the two alleles 42 s apart for each of the three consecutive separations. This technique shows potential to increase sample throughput in STR analysis techniques although separation resolution still needs to be improved.

摘要

已使用一种独特的样品引入和分离技术,对来自HUMVWF基因座的聚合酶链反应(PCR)扩增短串联重复序列(STR)样本进行了分析。使用一根毛细管将样品转移到超薄平板凝胶(57微米厚)上。这种超薄非变性聚丙烯酰胺凝胶用于分离扩增片段,并用溴化乙锭进行激光诱导荧光检测。通过检查重复样品的重现性,研究了使用该系统进行STR分析的可行性。通过比较14和17个HUMVWF等位基因在超薄平板凝胶上连续三次分离时的迁移情况来检验重现性。使用一个基因座,在连续三次分离中,每次分离时两个等位基因的迁移时间匹配,间隔为42秒。尽管分离分辨率仍需提高,但该技术显示出在STR分析技术中提高样品通量的潜力。

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