Step 1: QC and prepare samples
Prepare exactly 20 ng/µL in a minimum of 20 µL per sample. The samples you submit should either be full plasmids or PCR amplicons in water. This concentration should be measured using Qubit and impurity checked with a NanoDrop looking at the 260/230 and 280/230 ratios. Please submit these ratios and the elution liquid in the submission form.

For quality control before submission, you are also encouraged to perform a restriction digest for your plasmids. For amplicons, you must run a gel and ensure there's only a single band of your expected size, with no lower molecular weight or primer dimer bands visible. Amplicons must also be cleaned up before submitting. Check in if in doubt. Our lab uses bead cleans to remove small fragments from PCR samples, contact Carolina and Ziyan if you would like a protocol/demo on how to do this. 

Step 2: Complete this sequencing request form and sample spreadsheet, with your details and the details of your samples (+ plasmids reference maps if available). 

Step 3: Samples will be collected from the drop-off points every Wednesday around noon. Drop off your labelled samples to: 

• Option 1: The Schwessinger Lab (Room 3.078 Linnaeus Building). There is a drawer labelled "Samples for sequencing" in -20°C Freezer #1.
• Option 2: The Sanger sample Drop-off room (2.043 JCSMR). Please leave samples in the freezer compartment inside the designated container and using the racks provided.

Step 4: Collect your results from the general OneDrive folder the following Monday (or Tuesday if there is a public holiday) for everyone to access their data. Note: For data generated prior 2025, please use this link. 

A quick guide to the plasmid sequencing results can be found under the documents section below. If you submit amplicons for sequencing, you will receive a zipped Fastq folder (this contains the basecalled sequencing reads). 

Samples will be sequenced using the Nanopore Rapid Barcoding Kit 96 V14 (SQK-RBK114.96), on PromethION R10.4.1 (FLO-PRO114M) flowcells, using the BRF's PromethION device.

This operation continues to run on a cost recovery basis in a collaborative effort.