Genomics Research Core

Sanger Sequencing and Fragment Length Analysis

You must be a registered facility user before submitting any samples for processing. If your laboratory is new to our services, please register here.

Overview

The Genomics Research Core accepts a variety of templates, including PCR products, plasmids, BACs, etc. One sequencing run typically produces 850-950 nucleotides of high quality (98.5% accuracy) sequence data. The Genomics Research Core Sanger Sequencing Facility, located at 3343 Forbes Avenue in Pittsburgh, PA And can be contacted at gpclseq@pitt.edu or (412) 383-9077.

The Facility offers four levels of sequencing service:

  • PCR Clean-up and Sequencing Reaction: Submit your PCR reaction for cleanup followed by sequencing reaction with your separately submitted primers. The Genomics Research Core will supply all materials for sequencing reactions
  • PCR Sequencing Reaction: Submit PCR template and primers ready for addition of BigDye Terminator sequencing premix. Users need to supply DNA and primers. Appropriate clean up of DNA is the responsibility of the investigator, but we are happy to give advice and assistance.
  • Plasmid or BAC Sequencing Reaction: Submit purified plasmid or BAC template with primers or request addition of our universal primers: M13F, M13R, SP6, T3, T7 promoter, T7 terminator, SP6, FastBacF or FastBacR. . On the iLab sample submission form, select the appropriate primer from the drop down list in the “To be added by Core” column next to the sample ID. If you are running a sample with more than one primer, you will need to make a separate entry in the sample table for each sample:primer combination. Appropriate clean up of DNA is the responsibility of the investigator, but we are happy to give advice and assistance.
  • Capillary Run: Submit your completed reactions, appropriately prepared for sequencing or genotyping reactions for analysis on our capillary sequencer.

Fragment Length Polymorphism Analysis

The Genomics Research Core also provides support for fragment analysis technologies (AFLP and STRP genotyping) via instrument data generation only. Genomics Research Core does not perform PCR processing for these applications. PCR products can be submitted for capillary run. Instrument output data will be provided ready for data analysis yielding final genotyping calls by the investigator. Please contact the Genomics Research Core to initiate a project.

 

Sample Preparation and Submission:
PCR Clean-up and Sequencing
PCR Sequencing Only
Plasmid or BAC Sequencing
Capillary Run Only
Genotyping

Sanger Sequencing Methods and Data Delivery
Tips for primer design
Sanger Sequencing Scheduling and Turn Around
Sanger Sequencing FAQs