Large scale library generation for high throughput sequencing.
BACKGROUND: Large efforts have recently been made to automate the sample preparation protocols for massively parallel sequencing in order to match the increasing instrument throughput. Still, the size selection through agarose gel electrophoresis separation is a labor-intensive bottleneck of these p...
| Main Authors: | , , |
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| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2011-01-01
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| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC3083417?pdf=render |
| _version_ | 1831668623516631040 |
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| author | Erik Borgström Sverker Lundin Joakim Lundeberg |
| author_facet | Erik Borgström Sverker Lundin Joakim Lundeberg |
| author_sort | Erik Borgström |
| collection | DOAJ |
| description | BACKGROUND: Large efforts have recently been made to automate the sample preparation protocols for massively parallel sequencing in order to match the increasing instrument throughput. Still, the size selection through agarose gel electrophoresis separation is a labor-intensive bottleneck of these protocols. METHODOLOGY/PRINCIPAL FINDINGS: In this study a method for automatic library preparation and size selection on a liquid handling robot is presented. The method utilizes selective precipitation of certain sizes of DNA molecules on to paramagnetic beads for cleanup and selection after standard enzymatic reactions. CONCLUSIONS/SIGNIFICANCE: The method is used to generate libraries for de novo and re-sequencing on the Illumina HiSeq 2000 instrument with a throughput of 12 samples per instrument in approximately 4 hours. The resulting output data show quality scores and pass filter rates comparable to manually prepared samples. The sample size distribution can be adjusted for each application, and are suitable for all high throughput DNA processing protocols seeking to control size intervals. |
| first_indexed | 2024-12-19T20:34:06Z |
| format | Article |
| id | doaj.art-aeb82e9547f9469e8d0a53245eddcbc2 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-19T20:34:06Z |
| publishDate | 2011-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-aeb82e9547f9469e8d0a53245eddcbc22022-12-21T20:06:36ZengPublic Library of Science (PLoS)PLoS ONE1932-62032011-01-0164e1911910.1371/journal.pone.0019119Large scale library generation for high throughput sequencing.Erik BorgströmSverker LundinJoakim LundebergBACKGROUND: Large efforts have recently been made to automate the sample preparation protocols for massively parallel sequencing in order to match the increasing instrument throughput. Still, the size selection through agarose gel electrophoresis separation is a labor-intensive bottleneck of these protocols. METHODOLOGY/PRINCIPAL FINDINGS: In this study a method for automatic library preparation and size selection on a liquid handling robot is presented. The method utilizes selective precipitation of certain sizes of DNA molecules on to paramagnetic beads for cleanup and selection after standard enzymatic reactions. CONCLUSIONS/SIGNIFICANCE: The method is used to generate libraries for de novo and re-sequencing on the Illumina HiSeq 2000 instrument with a throughput of 12 samples per instrument in approximately 4 hours. The resulting output data show quality scores and pass filter rates comparable to manually prepared samples. The sample size distribution can be adjusted for each application, and are suitable for all high throughput DNA processing protocols seeking to control size intervals.http://europepmc.org/articles/PMC3083417?pdf=render |
| spellingShingle | Erik Borgström Sverker Lundin Joakim Lundeberg Large scale library generation for high throughput sequencing. PLoS ONE |
| title | Large scale library generation for high throughput sequencing. |
| title_full | Large scale library generation for high throughput sequencing. |
| title_fullStr | Large scale library generation for high throughput sequencing. |
| title_full_unstemmed | Large scale library generation for high throughput sequencing. |
| title_short | Large scale library generation for high throughput sequencing. |
| title_sort | large scale library generation for high throughput sequencing |
| url | http://europepmc.org/articles/PMC3083417?pdf=render |
| work_keys_str_mv | AT erikborgstrom largescalelibrarygenerationforhighthroughputsequencing AT sverkerlundin largescalelibrarygenerationforhighthroughputsequencing AT joakimlundeberg largescalelibrarygenerationforhighthroughputsequencing |