Submissions

General Instructions

  • All submissions to the HTSF should be made through the web-based submission system TracSeq (however if you are submitting samples for sequencing with PacBio and Ion Torrent systems, please continue to follow the specific submission information found on their respective pages).
  • Complete an electronic Study Initiation Form and then submit it through TracSeq.

  • Submitting samples to TracSeq requires an ONYEN account and password (follow the link for more information and to register).
  • Physical samples can be dropped off at 1153 Genome Sciences Building (map) from Mon-Fri between 10am and 4pm only please (unless otherwise arranged).  Due to UNC security considerations, our facilities may not be accessible outside of these hours.

  • All samples: All input amounts are assumed to be measured by a flourometer (e.g. Qubit). If using a spectrophotometer (e.g. Nanodrop) please compensate for overestimation and double the input amount.

How to submit a sample

  1. Please read over this entire page to get an idea about the process of preparing and submitting your sample(s).
  2. Contact Amy Perou, Corbin Jones, or Piotr MieczKowski regarding your planned submission(s).
  3. Download and fill out the appropriate submission form (PDF).
  4. Bring completed submission form(s) along with your DNA samples to the HTSF laboratory at 1153 GSB (map). 
  5. Open the freezer labelled "HTSF sample submission" and place samples in the yellow box marked "incoming samples".
  6. Using the information you provide with your submission form, the UNC Center for Bioinformatics will notify you when your data is available for download.

DNA Samples

A.   Standard DNA samples processing (TruSeq)

1.   Input Amount: 2 µg* of DNA
2.   Volume: 60 µl*

            * If you are unable to acquire the amount above, we can
              make libraries 
from smaller amounts of DNA but the
              probability of failure during the processing is higher.

B.   Low DNA samples (e.g. Chip-Seq)

1.    Input Amount: 10ng
2.    Volume: 35 µl 

C.   Amplicons

Please tell us the size(s)!  Standard library fragment sizes are usually 300 bp.

a.    100-600 – bp no fragmentation needed.
b.    >600bp – check the delicate fragmentation box. 

D.   cDNA and Fragmented DNA

1.   Standard library fragment sizes are usually 300 bp.  It is possible to make libraries from 100-600 bp,
     but if most of your fragments are larger than 1 kb, further fragmentation is required.

2.   Shearing according to the Illumina protocol for Covaris or BioRuptor gives sizes 200-400 bp.

a.   < 1kb – no fragmentation need, but please tell us the size(s).
b.   > 1kb – check the delicate fragmentation box
.

RNA Samples

A.      Standard RNA sample processing (TruSeq)

1.       Total RNA:

a.      Input Amount: 2 µg
b.      Volume: 50 µl

2.       mRNA (non-fragmented):

a.      Input Amount: 20 ng (purified from 1µg Total RNA)
b.      Volume: 10 µl

3.       Other: fragmented RNA or special request please contact us

B.      Small RNA samples (e.g. Micro-RNA)

1.       Total RNA:

a.      Input Amount: 2 µg
b.      Volume: 6 µl

2.       Purified small RNA

a.       Input Amount: 20 ng (purified from 1µg Total RNA)
b.       Volume: 6 µl
c.       Must be in molecular grade water or 10mM Tris-HCL, pH8.5. 

C.      Low RNA

1.       20ng – 100ng: Ambion MessageAmp premier RNA Amplification Kit

a.       Volume: 6 µl

2.       100pg – 10ng: Clontech SMARTer Ultra Low RNA kit for Illumina Seqencing

a.       Volume: 5 µl
b.       Please contact us
 if you wish to submit a sample of RNA that is this volume or smaller.

Libraries

A.      Required information with your sample submission

1.       Concentration
2.       Average library size
3.       Index number *
       
        * For multiplexing samples, each sample should have a unique index within each pool.  
          We ask that you also provide us with the identification of the index sequences used for
          each of the samples
.

B.      Pooled libraries for multiplexing

1.       Final concentration should be 15nM for each sample.
2.       Insufficient material?  Contact us
about pooling with lower concentrations.

PacBio Submissions

For more specific information and submission guidelines for PacBio sequencing, please see the HTSF's PacBio page.

Ion Torrent Submissions

For more specific information and submission guidelines for Ion Torrent sequencing, please see the HTSF's Ion Torrent page.



Additional notes

If you wish for more than one protocol to be performed (for different sizes or on different machines – e.g. IonTorrent) please contact us for additional information.