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Wild-type Blocking PCR Combined with Direct Sequencing as a Highly Sensitive Method for Detection of Low-Frequency Somatic Mutations
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Genetics
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JoVE Journal Genetics
Wild-type Blocking PCR Combined with Direct Sequencing as a Highly Sensitive Method for Detection of Low-Frequency Somatic Mutations

Wild-type Blocking PCR Combined with Direct Sequencing as a Highly Sensitive Method for Detection of Low-Frequency Somatic Mutations

DOI:
10.3791/55130-v

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10:41 min

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March 29, 2017

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, ,
1NeoGenomics Laboratories

Chapters

  • 00:05Title
  • 00:43Wild-Type Blocking PCR: Oligonucleotide Design
  • 03:54Purification of PCR Product by Magnetic Beads
  • 05:53Purification of the Sequencing Products
  • 08:17Identification of MYD88 Mutations in Human Samples
  • 10:03Conclusion

Summary

Automatic Translation

Automatic Translation

Wild-type blocking PCR followed by direct sequencing offers a highly sensitive method of detection for low frequency somatic mutations in a variety of sample types.

Tags

Keyword Extraction:Wild-type Blocking PCRSomatic MutationsMyd88 GeneBone Marrow SamplesExon FiveL265 HotspotM13 SequenceBlocking OligonucleotideWild-type TemplateMelting TemperatureExtension TemperatureSecondary StructureSelf-dimerization

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