Preparing samples for NGS
Yogendra Verma, Hanqin Li, Dirk Hockemeyer, Frank Soldner
Published: 2022-09-06 DOI: 10.17504/protocols.io.b4nwqvfe
Abstract
This protocol describes a standard procedure used to prepare PCR samples for Next Generation Sequencing (NGS)
Steps
1.
Run 5 µl PCR product in a 1% agarose gel to confirm successful amplification
2.
Purify the rest of the PCR product using column based or SPRI-beads based method, like Wizard SV Gel and PCR clean-up system or AMPure XP beads
3.
Following the instruction of the NGS facility, dilute the purified amplicon to a proper concentration, usually 10 ng/µl
4.
Send the purified samples to a NGS facility for multiplexing, library prep and NGS.
