Ultra-Low Coverage Sequencing as the Most Accurate Library Quantification Method Prior to Target Sequencing
- Авторлар: Krasnenko A.Y.1,2,3, Stetsenko I.F.1,3, Klimchuk O.I.1, Demkin V.V.4, Rakitko A.S.1,3,5, Surkova E.I.1, Druzhilovskaya O.S.3
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Мекемелер:
- Genotek Ltd.
- Pirogov Russian National Research Medical University
- Vavilov Institute of General Genetics
- Institute of Molecular Genetics
- Moscow State University, Faculty of Mechanics and Mathematics
- Шығарылым: Том 34, № 2 (2019)
- Беттер: 118-123
- Бөлім: Experimental Works
- URL: https://journals.rcsi.science/0891-4168/article/view/178356
- DOI: https://doi.org/10.3103/S089141681902006X
- ID: 178356
Дәйексөз келтіру
Аннотация
Accurate library quantification is very important during post-pooling captured target sequencing. There are a number of methods available to quantify libraries prior to sequencing, but no gold standard for the quantification of libraries exists. In this study, we compared common library quantification methods (Labchip, Qubit 3.0, qPCR with three primer sets) with ultra-low coverage sequencing (MiSeq with and without insert size correction). Cost, time and quantification accuracy were considered. We found that Qubit and MiSeq were better than qPCR and LabChip at predicting the final concentration. Also we revealed that MiSeq with insert size correction was the most accurate method for library quantification prior to target sequencing. This method allows for correction shifts in the ratio due to enrichment. Ultra-low coverage sequencing by Illumina MiSeq is the most accurate method for library quantification prior to pooling and post-pooling target enrichment.
Негізгі сөздер
Авторлар туралы
A. Krasnenko
Genotek Ltd.; Pirogov Russian National Research Medical University; Vavilov Institute of General Genetics
Email: esurkova@genotek.ru
Ресей, Moscow, 105120; Moscow, 117997; Moscow, 119333
I. Stetsenko
Genotek Ltd.; Vavilov Institute of General Genetics
Email: esurkova@genotek.ru
Ресей, Moscow, 105120; Moscow, 119333
O. Klimchuk
Genotek Ltd.
Email: esurkova@genotek.ru
Ресей, Moscow, 105120
V. Demkin
Institute of Molecular Genetics
Email: esurkova@genotek.ru
Ресей, Moscow, 123182
A. Rakitko
Genotek Ltd.; Vavilov Institute of General Genetics; Moscow State University, Faculty of Mechanics and Mathematics
Email: esurkova@genotek.ru
Ресей, Moscow, 105120; Moscow, 119333; Moscow, 119991
E. Surkova
Genotek Ltd.
Хат алмасуға жауапты Автор.
Email: esurkova@genotek.ru
Ресей, Moscow, 105120
O. Druzhilovskaya
Vavilov Institute of General Genetics
Email: esurkova@genotek.ru
Ресей, Moscow, 119333
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