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Capillary DNA sequencing and microsatellite analysis

eagle-i ID

http://eagle-i.itmat.upenn.edu/i/0000013f-2fa3-2a1e-5c87-8bc580000000

Resource Type

  1. Material analysis service

Properties

  1. Fee for service
    Yes
  2. Resource Description
    "The facility uses an ABI 3130xl capillary machine for Sanger-type sequencing with fluorescently-labeled dye-terminators, the ABI four-color detection, and sequence analysis system." "Purified DNA is provided by the investigator with an order form specifying the quantity, form (double-stranded, single-stranded, or PCR product) and estimated size of the DNA. It is essential that the size and concentration of PCR products are determined accurately prior to sequencing. Lambda or Bac clones require submission of greater quantities of DNA." "For each sequencing run, investigators must provide in numbered or labeled 1.5 ml Eppendorf tubes: <b>Double-stranded DNA:</b> 1-1.5 µg of purified DNA at 0.2-0.25 µg/µl <b>Single-stranded DNA:</b> 1 µg of purified DNA at 0.1-0.125 µg/µl <b>PCR products:</b> at least 15ng of purified DNA per 100 bp of PCR product in 5ul water <b>Lambda phage or Bac clones:</b> 10ug purified DNA at 2-2.5 µg/µl A name designation of 8 or fewer characters and the primers to be used for each run must be clearly indicated on the submission form. <b>M13 forward, M13 reverse, T7, T3, and SP6 primers</b> are provided by the Facility at no additional cost. Specific primers must be supplied by the investigator at 3.2 pmoles/µl. <b>Microsatellite Analysis: at least 450ng of DNA at 30ng/ul for 5 loci assay, and 10ng of DNA at 0.125ng/ul for 16 loci assay.</b> Depending on well availability, DNA samples delivered by 11:00 am will be available the next day at 3:00 pm." "For clean double-stranded templates, extremely accurate sequence reads of >=600 bases are obtained routinely. Text files of sequence data are e-mailed. The original result folder from the ABI base caller is placed in a computer folder for electronic retrieval if needed. Other modes of delivery can be arranged."
  3. Service Fee URL
    http://www.wistar.org/our-science/shared-facilities/genomics-facility/user-fees
  4. Contact
    Nguyen, Tran, M.S.
  5. Contact
    Capillary sequencing and microsatellite analysis specialist
  6. Related Resource
    ABI 3130xl genetic analyzer
  7. Service Provided by
    Genomics Facility (Wistar)
  8. Related Technique
    Microsatellite analysis
  9. Related Technique
    DNA sequencing assay
 
RDFRDF
 
Provenance Metadata About This Resource Record
  1. workflow state
    Published
  2. contributor
    fcoldren
  3. created
    2013-06-10T15:50:57.277-04:00
  4. creator
    fcoldren
  5. modified
    2013-07-02T10:27:29.371-04:00

Copyright © 2016 by the President and Fellows of Harvard College
The eagle-i Consortium is supported by NIH Grant #5U24RR029825-02 / Copyright 2016