Project submission process
1. Quote request form
Download, complete and send to igf@imperial.ac.uk.
2. Quotation
Sign and return the quotation received (ask for any changes needed before signing).
3. Sample submission form
Complete and return prior to submitting the samples to the Facility. Information on the “Container ID” column must exactly match the labels on your individual tubes/plates.
If you requested data analysis, we will need you to provide additional information regarding the experimental design, metadata and sample grouping. For this quote, we assume that each sample is only used once for data analysis, should any sample be needed more than once (e.g. control vs group1 and control vs group2) the cost for analysis will be adjusted by counting the control sample(s) twice.
4. Sample drop-off
- Prepare and deliver: samples/libraries to the facility in accordance with Sample Submission Guidelines (Further information below).
- QC Plate: a separate labelled QC plate is required with at least 4μl of each sample.
- Containers: samples/libraries must be submitted in 96-well plates, with samples loaded in columns.
- Label: plates with the Container ID, Quotation ID, the PI name and the date.
- An individual container ID needed on the form per each submitted plate.
- We cannot accept samples/libraries/data prior to the receipt of a sample submission form.
5. Project starts
- Initial QC: Your samples/libraries/data will be initially quality checked (QC) to make sure that the quality is acceptable for the requested application/service.
- Invoice: You will be invoiced according to the agreed quotation or any approved changes to your project.
- Result: You will receive results once your samples/libraries/data have been processed and the invoice has been paid.
Sample submission guidelines
Quantify libraries by qPCR and adjust the concentration of library or pool to 4-10nM by using Tris-Cl 10mM, pH 8.5 with 0.1% Tween 20.
Single library per lane:
- Submit 15 uL of 4-10 nM library
More than one library per lane(s):
- Submit 15 uL of 4-10 nM final pool per lane of sequencing requested
Prior to pooling barcoded libraries, it is essential to normalize the molar concentration of the libraries to ensure that an equal number of reads is generated for each library. In order to Convert ng/µl to nM and normalize library concentrations for pooling:
1. Determine the concentration (ng/µl) of each individual library using Qubit or KAPA Library Quantification Kits.
2. Determine the average size (bp) of each individual library using an Agilent Technologies 2100 Bioanalyzer.
3. Determine the molarity (nM) of each individual library using the figures above and the formula below:
4. Plan your library normalization (dilution calculations): Determine a common concentration (4-10nM) in order to dilute all individual libraries. Dilutions should be done using Tris-Cl 10mM, pH 8.5 with 0.1% Tween 20. Calculate the dilution of each individual library using the equation below:
5. Pool the normalized libraries: Combine equal volumes of each normalized library into a 1.5ml Eppendorf tube and gently pipette contents up and down 10 times to mix thoroughly. submit 15 µL of the pool per lane of sequencing requested.
6. The tube labels must exactly match the “Container ID” that you enter on the sample submission form. An individual ID must per provided per each tube or plate.
*Note: If the library pool concentration is below the submission concentrations mentioned above, contact IGF Facility for further instructions and advice.
Custom sequencing primers
If using custom sequencing primers, these must be submitted along with samples.
- Aliquots must be at 100 uM in at least 20 ul
- Primers must be diluted in EB or DI Water
- Must be in a Low-bind tube and clearly labelled
Important
- All libraries will be stored by the IGF Facility only for three months and then disposed of.
- Please contact us if you wish to collect your library after sequencing. We recommend submission of only an aliquote of your total library prep.
- Libraries received without the appropriate and correct paperwork or poorly labelled, will delay the initiation of your project, risk the safety of your samples and incur additional charges.