stilPCR increases the effective sequencing length of Illumina targeted next-generation sequencing.
Identifying pathogens, resistance-conferring mutations, and strain types through targeted amplicon sequencing is an important tool. However, due to the limitations of short read sequencing, many applications require the division of limited clinical samples. Here, we present stilPCR (single-tube Illu...
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| Main Authors: | , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2024-01-01
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| Series: | PLoS ONE |
| Online Access: | https://doi.org/10.1371/journal.pone.0314265 |
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| _version_ | 1841555382353264640 |
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| author | Jason D Limberis Roland J Nagel Soumitesh Chakravorty Dena Block Scott Dewell Alina Nalyvayko Zach Howard Grant Theron Rouxjeane Venter John Z Metcalfe |
| author_facet | Jason D Limberis Roland J Nagel Soumitesh Chakravorty Dena Block Scott Dewell Alina Nalyvayko Zach Howard Grant Theron Rouxjeane Venter John Z Metcalfe |
| author_sort | Jason D Limberis |
| collection | DOAJ |
| description | Identifying pathogens, resistance-conferring mutations, and strain types through targeted amplicon sequencing is an important tool. However, due to the limitations of short read sequencing, many applications require the division of limited clinical samples. Here, we present stilPCR (single-tube Illumina long read PCR), which allows the generation of hemi-nested amplicons in a single tube, with Illumina indexes and adapters, effectively increasing the Illumina read length without increasing the input requirements of reagents or sample. We have successfully utilized stilPCR on clinical sputum from tuberculosis patients to detect drug resistance mutations. |
| format | Article |
| id | doaj-art-518891a51ca44ccaba20bd4877470f6a |
| institution | Kabale University |
| issn | 1932-6203 |
| language | English |
| publishDate | 2024-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj-art-518891a51ca44ccaba20bd4877470f6a2025-01-08T05:33:28ZengPublic Library of Science (PLoS)PLoS ONE1932-62032024-01-011912e031426510.1371/journal.pone.0314265stilPCR increases the effective sequencing length of Illumina targeted next-generation sequencing.Jason D LimberisRoland J NagelSoumitesh ChakravortyDena BlockScott DewellAlina NalyvaykoZach HowardGrant TheronRouxjeane VenterJohn Z MetcalfeIdentifying pathogens, resistance-conferring mutations, and strain types through targeted amplicon sequencing is an important tool. However, due to the limitations of short read sequencing, many applications require the division of limited clinical samples. Here, we present stilPCR (single-tube Illumina long read PCR), which allows the generation of hemi-nested amplicons in a single tube, with Illumina indexes and adapters, effectively increasing the Illumina read length without increasing the input requirements of reagents or sample. We have successfully utilized stilPCR on clinical sputum from tuberculosis patients to detect drug resistance mutations.https://doi.org/10.1371/journal.pone.0314265 |
| spellingShingle | Jason D Limberis Roland J Nagel Soumitesh Chakravorty Dena Block Scott Dewell Alina Nalyvayko Zach Howard Grant Theron Rouxjeane Venter John Z Metcalfe stilPCR increases the effective sequencing length of Illumina targeted next-generation sequencing. PLoS ONE |
| title | stilPCR increases the effective sequencing length of Illumina targeted next-generation sequencing. |
| title_full | stilPCR increases the effective sequencing length of Illumina targeted next-generation sequencing. |
| title_fullStr | stilPCR increases the effective sequencing length of Illumina targeted next-generation sequencing. |
| title_full_unstemmed | stilPCR increases the effective sequencing length of Illumina targeted next-generation sequencing. |
| title_short | stilPCR increases the effective sequencing length of Illumina targeted next-generation sequencing. |
| title_sort | stilpcr increases the effective sequencing length of illumina targeted next generation sequencing |
| url | https://doi.org/10.1371/journal.pone.0314265 |
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