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
An usefulness of silica milk made with waste ultraviolet light tube for recovery of DNA fragment from agarose gel was represented. The glass milk is a water suspension of 50% fine silica powder prepared by grinding th...An usefulness of silica milk made with waste ultraviolet light tube for recovery of DNA fragment from agarose gel was represented. The glass milk is a water suspension of 50% fine silica powder prepared by grinding the crushed waste ultraviolet light tube with a porcelain mortar. It was showed that one microliter of the glass milk could bind more than 1 μg of DNA fragment, and DNA fragment in length from 125 bp to 23 kb could be efficiently recovered from agarose gel. The bound DNA could be eluted from the particle of SiO2 in the glass milk with a yield of about 60%–80%. The eluted DNA could be used in all manipulations in molecular cloning.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An usefulness of silica milk made with waste ultraviolet light tube for recovery of DNA fragment from agarose gel was represented. The glass milk is a water suspension of 50% fine silica powder prepared by grinding the crushed waste ultraviolet light tube with a porcelain mortar. It was showed that one microliter of the glass milk could bind more than 1 μg of DNA fragment, and DNA fragment in length from 125 bp to 23 kb could be efficiently recovered from agarose gel. The bound DNA could be eluted from the particle of SiO2 in the glass milk with a yield of about 60%–80%. The eluted DNA could be used in all manipulations in molecular cloning.
