Nanopore-only Microbial Isolate Sequencing Solution (NO-MISS) associated extraction protocols
Extraction Method Group
Nanopore-only Microbial Isolate Sequencing Solution (NO-MISS) associated extraction protocols
FOR RESEARCH USE ONLY
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) from cell cultures using SQK-RBK114 (.24 or .96) end-to-end protocol.
What extraction method is right for me?
| 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.
