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Recycling of superfine resolution agarose gel

Genetic markers are now routinely used in a wide range of applications, from forensic DNA analysis to marker-assisted plant and animal breeding. The usual practice in such work is to extract the DNA, prime the markers of interest, and sift them out by electrically driving them through an appropriate...

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Main Authors: Seng, Tzer Ying, Singh, R., Qamaruz Zaman, Faridah, Tan, Soon Guan, Syed Alwee, Sharifah Shahrul Rabiah
Format: Article
Language:English
English
Published: Fundacao de Pesquisas Cientificas de Ribeirao Preto 2013
Online Access:http://psasir.upm.edu.my/id/eprint/30339/1/Recycling%20of%20superfine%20resolution%20agarose%20gel.pdf
http://psasir.upm.edu.my/id/eprint/30339/
http://www.geneticsmr.com/issue/12/3?page=6
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spelling my.upm.eprints.303392016-01-28T08:17:27Z http://psasir.upm.edu.my/id/eprint/30339/ Recycling of superfine resolution agarose gel Seng, Tzer Ying Singh, R. Qamaruz Zaman, Faridah Tan, Soon Guan Syed Alwee, Sharifah Shahrul Rabiah Genetic markers are now routinely used in a wide range of applications, from forensic DNA analysis to marker-assisted plant and animal breeding. The usual practice in such work is to extract the DNA, prime the markers of interest, and sift them out by electrically driving them through an appropriate matrix, usually a gel. The gels, made from polyacrylamide or agarose, are of high cost, limiting their greater applications in molecular marker work, especially in developing countries where such technology has great potential. Trials using superfine resolution (SFR) agarose for SSR marker screening showed that it is capable of resolving SSR loci and can be reused up to 14 times, thus greatly reducing the cost of each gel run. Furthermore, for certain applications, low concentrations of agarose sufficed and switching to lithium borate buffer, instead of the conventional Tris-borate-ethylenediaminetetraacetic acid buffer, will further save time and cost. The 2.5% gel was prepared following the Agarose SFR™ manual by adding 2.5 g agarose powder into 100 mL 1X lithium borate buffer in a 250-mL flask with rapid stirring. Two midigels (105 x 83 mm, 17 wells) or 4 minigels (50 x 83 mm, 8 wells), 4 mm thickness can be prepared from 100 mL gel solution. A total of 1680 PCR products amplified using 140 SSR markers from oil palm DNA samples were tested in this study using SFR recycled gel. As average, the gel can be recycled 8 times with good resolution, but can be recycled up to 14 times before the resolutions get blurred. Fundacao de Pesquisas Cientificas de Ribeirao Preto 2013-03-11 Article PeerReviewed application/pdf en http://psasir.upm.edu.my/id/eprint/30339/1/Recycling%20of%20superfine%20resolution%20agarose%20gel.pdf Seng, Tzer Ying and Singh, R. and Qamaruz Zaman, Faridah and Tan, Soon Guan and Syed Alwee, Sharifah Shahrul Rabiah (2013) Recycling of superfine resolution agarose gel. Genetics and Molecular Research, 12 (3). pp. 2360-2367. ISSN 1676-5680 http://www.geneticsmr.com/issue/12/3?page=6 10.4238/2013.March.11.1 English
institution Universiti Putra Malaysia
building UPM Library
collection Institutional Repository
continent Asia
country Malaysia
content_provider Universiti Putra Malaysia
content_source UPM Institutional Repository
url_provider http://psasir.upm.edu.my/
language English
English
description Genetic markers are now routinely used in a wide range of applications, from forensic DNA analysis to marker-assisted plant and animal breeding. The usual practice in such work is to extract the DNA, prime the markers of interest, and sift them out by electrically driving them through an appropriate matrix, usually a gel. The gels, made from polyacrylamide or agarose, are of high cost, limiting their greater applications in molecular marker work, especially in developing countries where such technology has great potential. Trials using superfine resolution (SFR) agarose for SSR marker screening showed that it is capable of resolving SSR loci and can be reused up to 14 times, thus greatly reducing the cost of each gel run. Furthermore, for certain applications, low concentrations of agarose sufficed and switching to lithium borate buffer, instead of the conventional Tris-borate-ethylenediaminetetraacetic acid buffer, will further save time and cost. The 2.5% gel was prepared following the Agarose SFR™ manual by adding 2.5 g agarose powder into 100 mL 1X lithium borate buffer in a 250-mL flask with rapid stirring. Two midigels (105 x 83 mm, 17 wells) or 4 minigels (50 x 83 mm, 8 wells), 4 mm thickness can be prepared from 100 mL gel solution. A total of 1680 PCR products amplified using 140 SSR markers from oil palm DNA samples were tested in this study using SFR recycled gel. As average, the gel can be recycled 8 times with good resolution, but can be recycled up to 14 times before the resolutions get blurred.
format Article
author Seng, Tzer Ying
Singh, R.
Qamaruz Zaman, Faridah
Tan, Soon Guan
Syed Alwee, Sharifah Shahrul Rabiah
spellingShingle Seng, Tzer Ying
Singh, R.
Qamaruz Zaman, Faridah
Tan, Soon Guan
Syed Alwee, Sharifah Shahrul Rabiah
Recycling of superfine resolution agarose gel
author_facet Seng, Tzer Ying
Singh, R.
Qamaruz Zaman, Faridah
Tan, Soon Guan
Syed Alwee, Sharifah Shahrul Rabiah
author_sort Seng, Tzer Ying
title Recycling of superfine resolution agarose gel
title_short Recycling of superfine resolution agarose gel
title_full Recycling of superfine resolution agarose gel
title_fullStr Recycling of superfine resolution agarose gel
title_full_unstemmed Recycling of superfine resolution agarose gel
title_sort recycling of superfine resolution agarose gel
publisher Fundacao de Pesquisas Cientificas de Ribeirao Preto
publishDate 2013
url http://psasir.upm.edu.my/id/eprint/30339/1/Recycling%20of%20superfine%20resolution%20agarose%20gel.pdf
http://psasir.upm.edu.my/id/eprint/30339/
http://www.geneticsmr.com/issue/12/3?page=6
_version_ 1643830029373669376
score 13.211869

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