Evaluating insect microbiomes with Oxford Nanopore sequencing

Lawrence S. Blumer, Laura Avila, Tiana Scott, Anna J. Zelaya, Christopher W. Beck & Nicole M. Gerardo

https://doi.org/10.37590/able.v46.sup3

Supplemental Materials

• · Sample Pooling Spreadsheet [xlsx]
  • The Sample Pooling Spreadsheet is a MS Excel file that is used to assist in preparing equimolar samples that are combined to prepare a DNA library for sequencing as described in Appendix F Nanopore Library Preparation and Sequencing. This spreadsheet shows how to calculate the volume needed from each sample library (amplicons of the 16S gene with attached barcodes amplified from each sample via PCR) in the libraries pooling step of the protocol. In this step, samples are combined in a single tube. This pooled library will then be cleaned with magnetic beads, pelleted, and resuspended in elution buffer. An aliquot of the pooled library is used to load 50 ng of DNA to the Oxford Nanopore Flongle flow cell. There also are constrains on the volume to be loaded in the flow cell. This spreadsheet walks you through all these calculations. We recommend that you “Save a Copy” of this spreadsheet before entering your own values in the blue shaded cells.