Illumina Library Submission

Genomics/Bioinformatics Seminar being offered in September 2024

Sept 27, 2024 - So you want to do a: PacBio RNAseq Experiment

 

In Order to Get Into the CAT Illumina Sequencing Queue

  1. Submit a sequencing request and access the submission form through iLabs.
  2. If you don't have an iLabs account already you can review the account creation guide and create an account.
  3. Request services, select the appropriate sequencing run configuration (see description of services), and complete the sequencing submission form.

'Tube Name' should be in the following six-character format (your initials + PI initials + 2-digit number).

Examples: EDJD01, MJPW02 do not use any extra punctuation, spaces, etc.

The average size of the library (in base pairs), as seen on the Bioanalyzer trace.

The desired %PhiX spike in

Sample names (only letters, numbers, dashes, and underscores) and corresponding barcode sequence or name (in upper case)

Upload the bioanalyzer trace of your sample.

Submitting samples/tubes to the CAT Core

  1. Sample Tube: For a single lane of Illumina sequencing put 30ul of your sample in a 1.5ml low bind tube at 5-10nM (this is around 1.5-3ng/ul). If your library is lower than 5nM, please increase the volume to 40ul. If your sample is greater than 20 nM please dilute it. For each additional sequencing lane, please add an additional 20ul of the library. Label the top of the sample tube with the 'Tube Name' AND label the side of the sample tube with the date.
  2. Custom primers: put 20ul of a 100uM aliquot of the custom primer in a 1.5ml tube. Label the primer tube with your 'Tube Name' + 'Primer'
  3. Drop off sample:

Bring the Sample Tube to the CAT and any Custom Primers (if needed), in the small freezer on the ground outside the CAT Core's double doors. This is near room S271 of Genentech Hall.

  1. Shipping samples:

Place 1.5ml tubes in a secondary container like a 50ml conical with kimwipes to prevent samples from jostling. Ship via UPS or FedEx to:

UCSF CAT Lab Attn: GH-S252

600 16th Street

San Francisco CA 94158

 

Referencing the CAT Core and Publications

When submitting manuscripts, please acknowledge the CAT by including the text: “Sequencing was performed at the UCSF CAT, supported by UCSF PBBR, RRP IMIA, and NIH 1S10OD028511-01 grants.” 

 

To aid our facility in future grant submissions, it helps to reference labs and publications that have benefited from our core please notify us of your achievement by emailing catcore [at] ucsf.edu the PMID number, lab, and the piece of equipment used in the paper.