| Popis: |
This protocol is for the preparation of libraries for 96 samples of SARS-CoV-2 using the New England Biolabs NEBNext®Fast DNA Library Prep Set for Ion Torrent™ kit, ARTIC primer pools, the Ion Express Barcodes, and sequencing through Ion Torrent. The library preparation is semi-automated with the open source Opentrons robot. The volumes, concentrations, cycling conditions, and Opentrons protocols have been optimized through a series of experiments and are currently being used at our lab on a weekly basis. The samples used for this protocol were extracted after being obtained from nasopharyngeal swabs, placed in viral transport medium, and mixed 1:1 with AVL buffer (Qiagen). It is advisable to perform a screening for the samples before starting the library preparation by RT-qPCR and select the samples that have Ct values below 30. |
