PCR Protocols pp 401-402 | Cite as

Technical Notes for Sequencing Difficult Templates

  • David Stirling
Part of the Methods in Molecular Biology™ book series (MIMB, volume 226)


There are a number of template types that are generally recognized as being difficult to sequence. These can include sequences with a high guanine-cytosine (G/C) content, sequences that are very rich in adenine/thymine (A/T), sequences with a marked secondary structure, or large regions of homopolymer. There is no one solution to these difficulties; however, there are a number of approaches that can be used to improve the quality of sequencing data obtained from each type of problem sequence.


Secondary Structure Melting Temperature Large Region High Melting Temperature Problem Sequence 
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Copyright information

© Humana Press Inc. 2003

Authors and Affiliations

  • David Stirling
    • 1
  1. 1.Department of HematologyRoyal Infirmary of EdinburghEdinburghUK

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