Rapid recovery of DNA from agarose gel slices by coupling electroelution with monolithic SPE

Yu, Shengbing; Yang, Shaolin; Zhou, Ping; Zhou, Ke; Wang, Jing; Chen, Xiangdong

doi:10.1002/elps.200800777

Cited by 5 publications

(1 citation statement)

References 28 publications

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“…The fabrication of Si-APTES hybrid monolithic column was carried out by using CTAB, water, and ethanol as the porogen [13,25,34] as listed in Supplementary Materials Table S1. Basic condition was chosen for this preparation condition (pH 8).…”

Section: Preparation Of Si-aptes Hybrid Monolithic Columnmentioning

confidence: 99%

One-pot Facile Preparation of Amino-functionalized Silica Hybrid Monoliths for Mixed-mode Chromatography

2019

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Silica hybrid monolithic columns were prepared using two precursors, in which organo-functionalized trialkoxysilanes are mixed with tetraalkoxysilanes. In this study, several types of amino-functionalized silica hybrid monolithic columns were prepared via single-step "one-pot" approach, and the amount of silica precursors, porogens, as well as the reaction conditions were optimized. The preparation was carried out by mixing the silica precursors, i.e. tetraethoxysilane (TEOS) or tetramethoxysilane (TMOS) with amino precursors such as aminopropyltrimethoxysilane (APTES), aminoethylaminopropyl-trimethoxysilane (AEAPTMS), and phenylaminopropyltrimethoxysilane (PAPTMS) in a porogenic solution. The chromatographic performance of these hybrid monolithic columns was optimized by investigating several parameters through the separation of inorganic anions (IO3 -, BrO3 -, Br -, NO2 -, NO3 -, I -, SCN -) and some polar compounds (thymine, thymidine, adenosine, adenine, uridine). Results showed that the silica hybrid monolithic columns could be operated at higher flow-rate that favors rapid separation. The run-to-run repeatability of Si-APTES and Si-PAPTMS hybrid monolithic columns were satisfactory with relative standard deviations (n = 5) of less than 8% for all the analyte anions.Amino-functionalized silica precursors was used in the preparation of hybrid monolithic column to reduce the preparation time via one-pot approach. The amine groups in the structure can provide weak anion exchange interaction and also hydrophilic interaction. The synthesized hybrid monolithic columns revealed good mechanical stability and good separation repeatability.

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Section: Preparation Of Si-aptes Hybrid Monolithic Columnmentioning

confidence: 99%

One-pot Facile Preparation of Amino-functionalized Silica Hybrid Monoliths for Mixed-mode Chromatography

2019

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Isolation of Xis Gen Fragment of λ Phage from Agarose Gel Using Magnetic Particles for Subsequent Enzymatic DNA Sequencing

et al. 2012

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A reassessment of several erstwhile methods for isolating DNA fragments from agarose gels

Gao

Zhang

et al. 2021

3 Biotech

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Molecular biology research often requires extraction of DNA fragments from agarose gels. In the past decades, there have been many methods developed for this purpose. Currently most researchers, especially novices, use commercial kits for this extraction, although these kits cost money and the procedures involved are not necessarily easier than some erstwhile methods. We herein reintroduce and reassess several simple and cost-free older methods. One method involves excising a slice of the gel containing the DNA fragment, followed by a thaw-and-freeze procedure to release the DNA from the gel slice into the gel-making buffer. The second method involves a dialysis tubing and requires electroelution of the DNA from the gel slice in the tubing. The third one is to centrifuge the gel slice to release the DNA. The fourth method requires electro-transfer of the DNA from the gel into a filter paper, while the fifth one includes either allowing the DNA in the slice to be dissolved into a buffer or dissolving the DNA-containing gel slice, followed by DNA precipitation with ethanol or isopropanol. The strengths and weaknesses of these methods are discussed to assist researchers in making their choice. We also point out that some of the end uses of the DNA fragment in the agarose gel may not actually require extraction of the DNA. For instance, a tiny DNA-containing gel block or filter paper can be directly used as the template in a nested or semi-nested polymerase chain reaction to preliminarily determine the identity of the DNA fragment.

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Rapid recovery of DNA from agarose gel slices by coupling electroelution with monolithic SPE

Cited by 5 publications

References 28 publications

One-pot Facile Preparation of Amino-functionalized Silica Hybrid Monoliths for Mixed-mode Chromatography

One-pot Facile Preparation of Amino-functionalized Silica Hybrid Monoliths for Mixed-mode Chromatography

Isolation of Xis Gen Fragment of λ Phage from Agarose Gel Using Magnetic Particles for Subsequent Enzymatic DNA Sequencing

A reassessment of several erstwhile methods for isolating DNA fragments from agarose gels

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