We are happy to sequence client prepared Illumina-compatible libraries. These libraries undergo a MiSeq QC run to ensure they are suitable for HiSeq sequencing.
- Libraries not made at the OGBF are run on the HiSeq at the client’s risk.
- Libraries must be compatible with the Read 1, Indexing, and Read 2 primers used by the current Illumina chemistry.
- Dilutions must be based on quantitation by fluorometry or qPCR, and average size and/or primer/adapter dimer contamination assessed by using a Bioanalyzer or equivalent, e.g. Fragment Analyser, TapeStation.
- For single libraries, please submit >20 uL of library at 10 nM concentration diluted with RSB or EB, i.e. 10 mM Tris pH8.5 (with or without 0.01% Tween-20) in 1.5 mL low-bind tubes.
- For multiplexed library pools, unless otherwise discussed with us, please submit >20 uL at 10 nM (i.e. do not submit individual unpooled libraries) in 1.5 mL low-bind tubes.
- If you cannot submit a 10 nM dilution, then please contact us to discuss.
Client QC requirements prior to library submission
- Bioanalyser trace of the library or pooled libraries.
- Details of the library preparation that may be relevant to sequencing efficiency and demultiplexing.
- Quantification must be by using qubit/picogreen or qPCR.
- Download our sample submission form.
- Fill in the "User-supplied libraries" section, adding columns if necessary, and email the form to us.
- Index sequences must be submitted in the correct orientation for sample sheet entry for demultiplexing.