A modified method for whole exome resequencing from minimal amounts of starting DNA.
Next generation DNA sequencing (NGS) technologies have revolutionized the pace at which whole genome and exome sequences can be generated. However, despite these advances, many of the methods for targeted resequencing, such as the generation of high-depth exome sequences, are somewhat limited by the...
Saved in:
| Main Authors: | , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2012-01-01
|
| Series: | PLoS ONE |
| Online Access: | https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0032617&type=printable |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1850224596255506432 |
|---|---|
| author | Iwanka Kozarewa Juan Manuel Rosa-Rosa Christopher P Wardell Brian A Walker Kerry Fenwick Ioannis Assiotis Costas Mitsopoulos Marketa Zvelebil Gareth J Morgan Alan Ashworth Christopher J Lord |
| author_facet | Iwanka Kozarewa Juan Manuel Rosa-Rosa Christopher P Wardell Brian A Walker Kerry Fenwick Ioannis Assiotis Costas Mitsopoulos Marketa Zvelebil Gareth J Morgan Alan Ashworth Christopher J Lord |
| author_sort | Iwanka Kozarewa |
| collection | DOAJ |
| description | Next generation DNA sequencing (NGS) technologies have revolutionized the pace at which whole genome and exome sequences can be generated. However, despite these advances, many of the methods for targeted resequencing, such as the generation of high-depth exome sequences, are somewhat limited by the relatively large amounts of starting DNA that are normally required. In the case of tumour analysis this is particularly pertinent as many tumour biopsies often return submicrogram quantities of DNA, especially when tumours are microdissected prior to analysis. Here, we present a method for exome capture and resequencing using as little as 50 ng of starting DNA. The sequencing libraries generated by this minimal starting amount (MSA-Cap) method generate datasets that are comparable to standard amount (SA) whole exome libraries that use three micrograms of starting DNA. This method, which can be performed in most laboratories using commonly available reagents, has the potential to enhance large scale profiling efforts such as the resequencing of tumour exomes. |
| format | Article |
| id | doaj-art-4990299967334bd2b847b2b716fc5096 |
| institution | OA Journals |
| issn | 1932-6203 |
| language | English |
| publishDate | 2012-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj-art-4990299967334bd2b847b2b716fc50962025-08-20T02:05:35ZengPublic Library of Science (PLoS)PLoS ONE1932-62032012-01-0173e3261710.1371/journal.pone.0032617A modified method for whole exome resequencing from minimal amounts of starting DNA.Iwanka KozarewaJuan Manuel Rosa-RosaChristopher P WardellBrian A WalkerKerry FenwickIoannis AssiotisCostas MitsopoulosMarketa ZvelebilGareth J MorganAlan AshworthChristopher J LordNext generation DNA sequencing (NGS) technologies have revolutionized the pace at which whole genome and exome sequences can be generated. However, despite these advances, many of the methods for targeted resequencing, such as the generation of high-depth exome sequences, are somewhat limited by the relatively large amounts of starting DNA that are normally required. In the case of tumour analysis this is particularly pertinent as many tumour biopsies often return submicrogram quantities of DNA, especially when tumours are microdissected prior to analysis. Here, we present a method for exome capture and resequencing using as little as 50 ng of starting DNA. The sequencing libraries generated by this minimal starting amount (MSA-Cap) method generate datasets that are comparable to standard amount (SA) whole exome libraries that use three micrograms of starting DNA. This method, which can be performed in most laboratories using commonly available reagents, has the potential to enhance large scale profiling efforts such as the resequencing of tumour exomes.https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0032617&type=printable |
| spellingShingle | Iwanka Kozarewa Juan Manuel Rosa-Rosa Christopher P Wardell Brian A Walker Kerry Fenwick Ioannis Assiotis Costas Mitsopoulos Marketa Zvelebil Gareth J Morgan Alan Ashworth Christopher J Lord A modified method for whole exome resequencing from minimal amounts of starting DNA. PLoS ONE |
| title | A modified method for whole exome resequencing from minimal amounts of starting DNA. |
| title_full | A modified method for whole exome resequencing from minimal amounts of starting DNA. |
| title_fullStr | A modified method for whole exome resequencing from minimal amounts of starting DNA. |
| title_full_unstemmed | A modified method for whole exome resequencing from minimal amounts of starting DNA. |
| title_short | A modified method for whole exome resequencing from minimal amounts of starting DNA. |
| title_sort | modified method for whole exome resequencing from minimal amounts of starting dna |
| url | https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0032617&type=printable |
| work_keys_str_mv | AT iwankakozarewa amodifiedmethodforwholeexomeresequencingfromminimalamountsofstartingdna AT juanmanuelrosarosa amodifiedmethodforwholeexomeresequencingfromminimalamountsofstartingdna AT christopherpwardell amodifiedmethodforwholeexomeresequencingfromminimalamountsofstartingdna AT brianawalker amodifiedmethodforwholeexomeresequencingfromminimalamountsofstartingdna AT kerryfenwick amodifiedmethodforwholeexomeresequencingfromminimalamountsofstartingdna AT ioannisassiotis amodifiedmethodforwholeexomeresequencingfromminimalamountsofstartingdna AT costasmitsopoulos amodifiedmethodforwholeexomeresequencingfromminimalamountsofstartingdna AT marketazvelebil amodifiedmethodforwholeexomeresequencingfromminimalamountsofstartingdna AT garethjmorgan amodifiedmethodforwholeexomeresequencingfromminimalamountsofstartingdna AT alanashworth amodifiedmethodforwholeexomeresequencingfromminimalamountsofstartingdna AT christopherjlord amodifiedmethodforwholeexomeresequencingfromminimalamountsofstartingdna AT iwankakozarewa modifiedmethodforwholeexomeresequencingfromminimalamountsofstartingdna AT juanmanuelrosarosa modifiedmethodforwholeexomeresequencingfromminimalamountsofstartingdna AT christopherpwardell modifiedmethodforwholeexomeresequencingfromminimalamountsofstartingdna AT brianawalker modifiedmethodforwholeexomeresequencingfromminimalamountsofstartingdna AT kerryfenwick modifiedmethodforwholeexomeresequencingfromminimalamountsofstartingdna AT ioannisassiotis modifiedmethodforwholeexomeresequencingfromminimalamountsofstartingdna AT costasmitsopoulos modifiedmethodforwholeexomeresequencingfromminimalamountsofstartingdna AT marketazvelebil modifiedmethodforwholeexomeresequencingfromminimalamountsofstartingdna AT garethjmorgan modifiedmethodforwholeexomeresequencingfromminimalamountsofstartingdna AT alanashworth modifiedmethodforwholeexomeresequencingfromminimalamountsofstartingdna AT christopherjlord modifiedmethodforwholeexomeresequencingfromminimalamountsofstartingdna |