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DNA Sequencing Variable Regions

Want to know more about your antibodies?  From mRNA to protein sequence…let our experts do the decoding! 

Our molecular biology services:

  • Messenger RNA isolation from hybridoma cells.
  • DNA sequencing of immunoglobulin heavy and light chain variable regions.
  • Edman Degradation for sequence confirmation.
  • Cloning of variable region DNA into vectors followed by transformation into competent cells. 

Benefits:

  • Sequencing can provide valuable insight on the structure of you antibody. 
  • Knowing the structure of your antibody can shed light on its interactions with other molecules.

Common applications:

  • Patents – documenting your antibody sequence helps to protect your discovery.
  • Differentiating antibodies made to a common target.

Technique:

At GMA, we employ a rigorous strategy to ensure that your antibody sequence is accurate!  We begin by isolating only cytoplasmic, processed RNA – free of genomic DNA contamination.  This is followed by a 5’ RACE method, allowing for amplification of the 5’ end of the variable region, which is traditionally difficult to prime.  This method, in combination with a set of nested PCR’s using gene specific primers extending from the constant region, results in complete variable region sequences.

Reliability:

All sequencing done at GMA has been crossed referenced with protein sequence obtained via Edman Degradation at an independent facility.  Sequences have matched 100%.  We also reference the guidelines provided by http://www.bioinf.org.uk/abs/ to locate and highlight the complementarity determining regions (CDR’s).  All sequences generated have contained the pattern of amino acids, or landmarks, most commonly surrounding the CDR’s.

Presentation:

Data is presented in both a nucleotide sequence and a protein sequence format. 

Your results will be emailed to you within 2-4 weeks.

Contact us.

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Copyright © 2009 Green Mountain Antibodies, Inc. All Rights Reserved
PO Box 1283, 1 Mill Street, Suite 1-7, Burlington, VT  05402 

p 802.865.6230   f 802.865-0115