Nanopore-only Microbial Isolate Sequencing Solution (NO-MISS) associated extraction protocols

We have developed four optimised extraction methods to generate high quality genomic DNA from your cell cultures, allowing maximised sequencing output using the Nanopore-only Microbial Isolate Sequencing Solution (NO-MISS) - Rapid Barcoding Kit V14 (SQK-RBK114.24 or SQK-RBK114.96) end-to-end protocol.

What extraction method is right for me?

Decision tree extract methods SVG


Sample extraction method Sample type Sample Input Expected yield Expected DNA Integrity Number (DIN) Average sequencing read lengths Extraction kits used
Universal bead-beating gDNA extraction Universal applications: bacteria, fungi or yeast 1 ml liquid overnight culture (~1 x 10^8 – 10^9 cfu/ml) or half of a loop of colonies from a plate >200 ng/µl per sample 7-9 ~4-7 kb QIAGEN PowerBead Tube and Promega Maxwell® RSC PureFood Pathogen kit
Bacteria gDNA extraction Bacterial 200 µl liquid overnight culture (~1 x 10^8 – 10^9 cfu/ml) or 1/8 of a loop of colonies from a plate ~15-20 ng/µl per sample 9 >7 kb - Size will vary based on sample input species NEB Monarch® Spin gDNA Extraction Kit
Hard to lyse organisms gDNA extraction Mycobacterium tuberculosis
(or hard to extract bacterial samples)
5 – 10 mg cells from solid or liquid media ~15-40 ng/µl per sample 8 >7 kb - Size will vary based on sample input species NEB Monarch Spin gDNA Extraction Kit
Fungi gDNA extraction Fungi or yeast 2 ml of ~1 x 10^7 cfu/ml overnight culture or a full 10 µl inoculating loop from a plate ~40 ng/µl per sample N/A >7 kb - Size will vary based on sample input species NEB Monarch Spin gDNA Extraction Kit

Note: The yield, DIN and sequencing read length of extracted DNA may vary depending on sample quality and species. Please ensure you are following the correct method and using high-quality sample inputs.