Services
– Columns, DNA Size-Select
• Columns for DNA Size-Selection
• Alternative DNA Size-Selection methods
• Online Submission Request
• Physical Submission of Samples
DNA Purification & Size-Selection
The Core uses the Zymo columns described below for rapid DNA size-selection. For more information, visit either the distributor's website (
Genesee Scientific) or the manufacturer's website (
Zymo Research).
Select-A-Size DNA Clean & Concentrator Kit — (Cat #D4080):
Size-Select DNA with retention points of ≥50-bp, ≥100-bp, ≥150-bp, ≥200-bp, ≥300-bp (other cutoffs between 50-300 bp can be attempted by titration).
Clean and concentrate up to 3 µg DNA in as little as 10-µl elution with 0-µl wash residue carryover.
Eluted DNA is of high quality and well suited for NGS, ligations, restriction endonuclease digestions, and library preparation cleanup.
DNA Size Limits: 50 bp to 23 kb.
Note 1: Maximum depletion of a lower-size fragment requires that the selected cutoff value be at least 50-bp higher than the unwanted fragment size; using this criterion, we have been extremely successful at eliminating adapter-dimers from NGS libraries while retaining the bulk of the desired fragment sizes.
Note 2: Conversely, maximum enrichment of desired fragments requires that the lower cut-off be at least 100-bp lower than the desired fragments; thus, there can be a tension between depletion of adapter-dimers and retention of the desired fraction of library if the difference in sizes is <100-bp. In that case, adjustments to the protocol can be attempted to find a happy 'medium' between the two objectives.
Note 3: Although we highly recommend the 'lower-cutoff' size-selection protocol with this kit, we recommend caution with respect to using the double-size selection protocol (i.e., lower cutoff = 300 bp; upper cutoff = 700-bp). Nevertheless, it should be noted that we have performed the double-size selection protocol on only a few libraries; thus, it is possible that the problems we encountered with that protocol were library-specific rather than any reflection on the functionality of the Select-A-Size DNA Clean & Concentrator Kit.
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Alternative DNA Size-Selection methods
Depending on your research needs, DNA size-selection by the BluePippin or E-gel might be more appropriate. For product specifications and other information regarding these platforms, please see the following web pages:
BluePippin
E-gel
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Online Submission Request
Log In to the website, and click on "Services Page":
- Make the necessary 'selections'.
- In the "Client Memo" field, specify the desired elution volume and any other information. [Note: Unless otherwise
requested, DNA will be eluted with TVLE (10 mM Tris, 0.05 mM EDTA).
- Click the 'check box', and submit your request.
If you make an error in your request, you may revise that request if you do so before we begin processing it.
You will receive an automated email when your samples are processed and ready for pick-up.
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Physical Submission of Samples
Tubes: Samples exceeding ~25 ul should be submitted in 0.5-1.5 ml tubes (standard or Eppendorf LoBind); otherwise, 0.2-ml tubes are acceptable.
Label Submission: Upon successful submission of your online request, the 'Submission #" will appear on your screen. Then, label the tube rack itself with:
- Submission #;
- Date;
- PI's Last Name; and,
- Submitter name or initials.
Delivery site: Transfer tubes to a Genomics Core rack (taking your own rack back to
your lab), and put samples in the Mini-fridge (by sink) in the Genomics Core (A628 Life Science Annex).
[Note: If the Core is locked, put your samples in the plastic "After-Hours" GC drop-box located in the adjacent Cold Room (beside sink, Rm. A650).]
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