3rd Dec 2020
Updated the "Notes on multiple crRNAs" in the Preparing the Cas9 ribonucleoprotein complexes (RNPs) step to say:
If you have validated a set of crRNA probes that are always run together, we recommend pooling all crRNA probes and then aliquoting that pool and freezing each aliquot at -80°C. Each time you run a Cas9 experiment and need to make an RNP complex, take out a crRNA pool aliquot and combine with the tracrRNA.
3rd Dec 2020
Updated the "Notes on multiple crRNAs" in the Preparing the Cas9 ribonucleoprotein complexes (RNPs) step to say:
If you have validated a set of crRNA probes that are always run together, we recommend pooling all crRNA probes and then aliquoting that pool and freezing each aliquot at -80°C. Each time you run a Cas9 experiment and need to make an RNP complex, take out a crRNA pool aliquot and combine with the tracrRNA.
20th September 2019
- A note has been added at the end of the library preparation instructions about using the Sequencing Auxiliary Vials (EXP-AUX001) for splitting a library into two aliquots
20th September 2019
- A note has been added at the end of the library preparation instructions about using the Sequencing Auxiliary Vials (EXP-AUX001) for splitting a library into two aliquots
17th September 2019
- The NEB Quick Dephosphorylation Kit has been discontinued; the new recommendation is to use Quick CIP (NEB, cat # M0525). This kit also contains the NEB CutSmart Buffer, so it is no longer necessary to purchase it separately.
17th September 2019
- The NEB Quick Dephosphorylation Kit has been discontinued; the new recommendation is to use Quick CIP (NEB, cat # M0525). This kit also contains the NEB CutSmart Buffer, so it is no longer necessary to purchase it separately.
23rd August 2019
- The protocol structure has been shortened and simplifed
- Information has been added about enrichment by 'tiling' a large genomics region
- Troubleshooting guide added, detailing most commonly-seen issues with nanopore sequencing, and how to resolve them
23rd August 2019
- The protocol structure has been shortened and simplifed
- Information has been added about enrichment by 'tiling' a large genomics region
- Troubleshooting guide added, detailing most commonly-seen issues with nanopore sequencing, and how to resolve them
Updated with new information for MinKNOW release v19.06:
- Compression of queued reads
- Pause functionality
- UI improvements including:
- Flow cell health at platform QC
- Flow cell status on landing page
- R10 flow cell running scripts
- New Fast Models for RNA and DNA
- PDF report and CSV files for Duty time and Cumulative output are now found in same location as actual reads and fastQ files
- Flongle hardware check for confirming Flongle adapter functions correctly
Update to the flow cell priming section, and instructions for topping up the flow cell with fuel, following the release of the Flow Cell Priming Kit (EXP-FLP002)