All sequencing samples must be submitted with accurate library base pair range information (Bioanalyzer/Tape Station/Fragment Analyzer results) or service must be requested per submitted sample. Gel images are not accepted for sequencing libraries only gDNA/RNA for prep. Please submit traces with the x-axis in bp NOT seconds. If possible please also place a smear range around your library and generate a molarity calculation with the analysis software. Label your samples clearly and with the same ids as indicated on submitted tube(see recommendations below).
Please also submit samples in water (or Tris-HCl) where possible, especially if submitting for the HiSeq4000/NovaSeq where a speedvac may be needed to concentrate samples to the required concentration for sequencing.
For Single Tube Sample Submission to the VC GSL we require the following:
- One signed copy of our current Sequencing Submission Form for the MiSeq sequencing, NovaSeq sequencing, or HiSeq4000 sequencing. Please email a pdf copy to email@example.com and include a hard-copy with the library submission.
- Copy of Bioanalyzer traces (PDF see notes above) sent to firstname.lastname@example.org for the components of/or the pooled submitted library (unless a Bioanalyzer trace is requested on the form).
- An Excel Sheet (not PDF) with Sample names and indexes listed emailed to email@example.com (only required if sample being submitted is multiplexed)
- Samples in 1.5mL low-binding tubes (~10uL of at least 10nM), normalization not required if concentration noted). Please note your quantification method and suspension buffer type on submission form. Extra qPCR will be charged for samples submitted at concentrations higher than 100nM without notification that a dilution may be necessary.
- Sample names labeled on the TOP and sides of the tubes (PI Initials, Your Initials, Number, ex: DRMC001, and month/year)
For Submitting Multiplexed libraries as separate tubes:
IMPORTANT TO READ!!!Please follow the above parameters and provide all of the same documentation but note that we charge for additional QC required to multiplex samples submitted in separate tubes. Each additional sample to be pooled (beyond the first sample per lane) will be charged a qPCR fee to ensure accurate pooling. If you have Qubit values, please also submit these to help ensure accurate pooling. A $500+ pooling test can be requested to ensure accurate pooling for large super-pool experiment. We do not guarantee any pooling accuracy but are generally dead-on and reasonable with redos where pooling was really off the mark unless dealing with hard to quantify samples (multiple peaks, very broad).
Also, please name subsequent sample with the same library base name (PI Initials, Your Initials, Number, ex: DRMC001) followed by a letter (A-Z) or (_01-_0XX). [DRMC001A, DRMC001B, DRMC002A, DRMC002B would be two lanes of samples with two samples per lane.
Please submit an excel spreadsheet with sample names, concentration, index sequence, and requested pooling percentages with your other required documents to firstname.lastname@example.org. If you do not specify on your form and in an excel that you want uneven pooling your samples will be pooled equimolar and check with the facility to make sure your pooling wishes are even possible.
The GSL is NEVER responsible for failures related to custom sequencing or indexing primers. The user is responsible for primers design. It is very obvious when a run fails due to an unannealed custom primer. Custom primers should be provided to the facility at 100um in 10-20ul aliquots. HiSeq4000 can only accommodate R1 custom primers.
Sample Drop off Locations:
Shana McDevitt/Dylan Smith
B206 Stanley Hall (Basement Level 2)
Shipping Samples to GSL (Sequencing Only, Library Prep goes to FGL)
Please ship your samples on dry ice overnight to the following address:
B206 Stanley Hall
Berkeley, CA 94720-3220
If you have any questions regarding sample submission, please contact Shana McDevitt, email@example.com or the lab directly firstname.lastname@example.org