Conventional methods of DNA recovery from agarose gel generally require expensive equipment, extended elution times, or considerable handling of the sample after elution. We developed a simple protocol for a quick and...Conventional methods of DNA recovery from agarose gel generally require expensive equipment, extended elution times, or considerable handling of the sample after elution. We developed a simple protocol for a quick and effective recovery of DNA from agarose gels with good yield and quality. Using a Sephadex resin filled spin column, DNA fragments of 500 bp to 6 kb in an agarose gel slice were easily recovered by a 2 min centrifugation. The recovery efficiencies were over 40% -50% and the eluted DNA can be used directly for downstream application, such as polymerase chain reactions （PCR） and restriction enzyme digestion. This method could also be used to recover large DNA fragment （48 kb） without degradation. The use of Sephadex helps to remove small molecular impurities from agarose and it also reduces the chance of clogging the column filter caused by direct contact with agarose.展开更多
本文以云南普洱熟茶为实验材料,以不同浓度乙醇水溶液为洗脱液,利用MCI GEL CHP 20P（75~150μm）树脂色谱柱和SephadexTMLH-20葡聚糖凝胶色谱柱,并结合高效液相色谱（HPLC）法,对普洱熟茶中主要黄酮醇类物质进行了分离和定性。结果表...本文以云南普洱熟茶为实验材料,以不同浓度乙醇水溶液为洗脱液,利用MCI GEL CHP 20P（75~150μm）树脂色谱柱和SephadexTMLH-20葡聚糖凝胶色谱柱,并结合高效液相色谱（HPLC）法,对普洱熟茶中主要黄酮醇类物质进行了分离和定性。结果表明：普洱熟茶提取液浓缩后过MCI GEL CHP 20P（75~150μm）树脂色谱柱,经不同浓度（10%,30%,50%,70%,90%）乙醇溶液洗脱分离后,将获得的70%和90%的洗脱液过SephadexTMLH-20葡聚糖凝胶色谱柱,70%的洗脱液经90%乙醇溶液洗脱分别获得了HPLC纯度为96.0%的杨梅素和HPLC纯度为98.5%的槲皮素;90%的洗脱液经70%的乙醇溶液洗脱获得了HPLC纯度为95.6%的山奈酚。展开更多
文摘Conventional methods of DNA recovery from agarose gel generally require expensive equipment, extended elution times, or considerable handling of the sample after elution. We developed a simple protocol for a quick and effective recovery of DNA from agarose gels with good yield and quality. Using a Sephadex resin filled spin column, DNA fragments of 500 bp to 6 kb in an agarose gel slice were easily recovered by a 2 min centrifugation. The recovery efficiencies were over 40% -50% and the eluted DNA can be used directly for downstream application, such as polymerase chain reactions （PCR） and restriction enzyme digestion. This method could also be used to recover large DNA fragment （48 kb） without degradation. The use of Sephadex helps to remove small molecular impurities from agarose and it also reduces the chance of clogging the column filter caused by direct contact with agarose.
文摘本文以云南普洱熟茶为实验材料,以不同浓度乙醇水溶液为洗脱液,利用MCI GEL CHP 20P（75~150μm）树脂色谱柱和SephadexTMLH-20葡聚糖凝胶色谱柱,并结合高效液相色谱（HPLC）法,对普洱熟茶中主要黄酮醇类物质进行了分离和定性。结果表明：普洱熟茶提取液浓缩后过MCI GEL CHP 20P（75~150μm）树脂色谱柱,经不同浓度（10%,30%,50%,70%,90%）乙醇溶液洗脱分离后,将获得的70%和90%的洗脱液过SephadexTMLH-20葡聚糖凝胶色谱柱,70%的洗脱液经90%乙醇溶液洗脱分别获得了HPLC纯度为96.0%的杨梅素和HPLC纯度为98.5%的槲皮素;90%的洗脱液经70%的乙醇溶液洗脱获得了HPLC纯度为95.6%的山奈酚。
