An efficient method for purification of PCR products for sequencing.

by: H Ma, S Difazio

BioTechniques, Vol. 44, No. 7. (June 2008), pp. 921-923.

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Abstract

A high-throughput DNA sequencing method that generated high quality data was developed. A frame fashioned from a standard agarose gel combined with 0.1%-0.2% low-melting point (LMP) agarose gel was used to isolate the PCR product of interest. Collected PCR products were centrifuged without any reagents and the supernatants were directly used for a sequencing reaction. This method is simple and labor efficient, provides high quality sequences at a low cost, and bypasses problems with impure PCR products. This technique has been used for single nucleotide polymorphism (SNP) discovery in Populus angustifolia trees.

@article{citeulike:3017876, abstract = {A high-throughput DNA sequencing method that generated high quality data was developed. A frame fashioned from a standard agarose gel combined with 0.1\%-0.2\% low-melting point (LMP) agarose gel was used to isolate the PCR product of interest. Collected PCR products were centrifuged without any reagents and the supernatants were directly used for a sequencing reaction. This method is simple and labor efficient, provides high quality sequences at a low cost, and bypasses problems with impure PCR products. This technique has been used for single nucleotide polymorphism (SNP) discovery in Populus angustifolia trees.}, address = {Department of Biology, West Virginia University, Morgantown, WV 26506, USA. hao.ma@mail.wvu.edu}, author = {Ma, H. and Difazio, S. }, citeulike-article-id = {3017876}, doi = {http://dx.doi.org/10.2144/000112809}, issn = {0736-6205}, journal = {BioTechniques}, keywords = {pcr, sequencing}, month = {June}, number = {7}, pages = {921--923}, posted-at = {2008-07-18 14:08:04}, priority = {2}, title = {An efficient method for purification of PCR products for sequencing.}, url = {http://dx.doi.org/10.2144/000112809}, volume = {44}, year = {2008} }

RIS record

TY - JOUR ID - citeulike:3017876 L3 - citeulike-article-id:3017876 TI - An efficient method for purification of PCR products for sequencing. JF - BioTechniques VL - 44 IS - 7 SP - 921 EP - 923 AD - Department of Biology, West Virginia University, Morgantown, WV 26506, USA. hao.ma@mail.wvu.edu SN - 0736-6205 N2 - A high-throughput DNA sequencing method that generated high quality data was developed. A frame fashioned from a standard agarose gel combined with 0.1%-0.2% low-melting point (LMP) agarose gel was used to isolate the PCR product of interest. Collected PCR products were centrifuged without any reagents and the supernatants were directly used for a sequencing reaction. This method is simple and labor efficient, provides high quality sequences at a low cost, and bypasses problems with impure PCR products. This technique has been used for single nucleotide polymorphism (SNP) discovery in Populus angustifolia trees. KW - pcr KW - sequencing AU - Ma, H AU - Difazio, S PY - 2008/06// UR - http://dx.doi.org/10.2144/000112809 ER -

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