General Submission Guidelines
- Please submit your order form before sending in samples.
- If you are submitting more than 24 samples, put the samples in a 96-well plate and include and Excel file with the sample layout in your order form.
- Clearly mark all sample submissions with your order number.
- Include any QC information (Qubit data, Bioanalyzer or Fragment Analyzer run, gel picture, etc) about your sample on your order form.
- DNA should have absorbance ratio values of 1.8 – 2.0, and be submitted in 1x TE.
- RNA samples should be suspended in RNase-free water, or 1x TE buffer prepared with RNAse-free water.
Libraries prepared at the GGBC can be sequenced on the Flongle or MinION Nanopore flow cells. If you have a project for Oxford Nanopore Sequencing and do not see a library preparation for your experiment, please contact us for custom sequencing.
| ||Flongle (FLO-FLG106)||MinION (FLO-MIN106)
|Yield per flow cell||1-2 Gb||15-30Gb for DNA
1-3 Gb for RNA
|Channels per flow cell||126||512
|Price per flow cell||$122*||$999*
|Run time||1 min - 16 hours||1 min - 48 hours
*see pricing tab for more information on costs
We offer library preparation for gDNA, amplicons, cDNA, and direct RNA. Not all library preparation types are compatible with both flow cells. See the chart below for details about each type.
|Library Prep |
|DNA Ligation ||Direct RNA||Rapid Ligation||Amplicon/ |
Amplicon/cDNA with barcoding
|500ng-1ug of dsDNA||500ng-1ug RNA||400ng HMW gDNA||100-200 fmol amplicons or cDNA||10ng gDNA||0.5nM amplicons/cDNA
|Shearing||optional Covaris shearing||none||yes||none||none||none
|Mulitplexed||no||no||no||no||yes, up to 12 samples||yes, up to 96 samples
|Flongle/MinION compatibility||compatible with both||not for Flongle currently||compatible with both||not for Flongle currently||not for Flongle currently||not for Flongle currently
The DNA Ligation library preparation is best for double-stranded DNA, with optional fragmentation. This protocol does not use PCR. This library prep will include a positive control strand to help distinguish between sample failure and library prep failure in troubleshooting. The control strand is a 3.6kb amplicon from the Enterobacteria phage Lambda genome. If you are using this protocol for long reads, please make sure to submit high-molecular weight DNA.
- Please submit 500ng-1ug of double-stranded DNA
The Direct RNA library preparation is for sequencing direct RNA sequencing. This protocol does not use PCR. Input RNA should have a 3’ polyA tail. On the MinION, this will yield 1-3 Gb, when starting with high-quality RNA.
- Please submit 500ng-1ug of RNA
The Rapid Ligation preparation is a shorter protocol for sequencing gDNA. The protocol does not use PCR, and uses transposase-based fragmentation. The read length will have a random distribution, depending on the input fragment size.
- Please submit 400ng of high-molecular weight gDNA
The Amplicon/cDNA library preparation is for sequencing shorter DNA fragments, such as amplicons and cDNA. This protocol does not use PCR. Currently, this preparation is only for single-samples, not multiplexing. Submit cDNA samples pre-made. For amplicons, you can either submit gDNA and your region-specific primer sequences and we will make the amplicons, or you can submit your amplicons directly. We will only make amplicons when the target size is <1,000 bp.
For pre-made amplicons and cDNA:
- Please submit 1-1.5ug of long double-stranded DNA, or 100-200 fmol for short fragments
For gDNA, to be made into amplicons by GGBC:
- Please submit 500ng-2ug of gDNA
- Please submit your primer sequences
The 16S library preparation is for sequencing the entire 16s rRNA region. This protocol uses PCR. Amplicons can be barcoded for up to 12 samples.
- Please submit 10ng of gDNA
Coming soon: Amplicon/cDNA with barcoding for 1-96 samples
The Amplicon/cDNA library preparation with barcoding is for sequencing multiple samples on the same flow cell. This protocol does use PCR, and can barcode up to 96 samples.