title: Locked Nucleic Acid Probes (LNA) for Enhanced Detection of Low-Level, Clinically Significant Mutations.
abstract: The detection of clinically significant somatic mutations present at low level in a tissue sample represents a challenge in any laboratory. While several high sensitivity methods are described, the incorporation of these new techniques in a clinical lab may be difficult if the technology is not readily available or requires major changes in the workflow of the laboratory. Techniques that are robust and easily adapted to existing laboratory protocols are highly advantageous. In this chapter we describe the use of locked nucleic acid (LNA) probes to modify existing polymerase chain reaction (PCR)-based protocols which can then be sequenced by Sanger sequencing. LNA probes are used to enhance the sensitivity of Sanger sequencing to mutation frequencies below 1 %. The method is robust and is easily incorporated for assessment of any sample with low tumor content or low mutant allele burden.
authors: Department of Pathology, Memorial Sloan Kettering Cancer Center, 1275 York Avenue, New York, NY, 10065, USA. NafaK@mskcc.org.
authors: Khedoudja
authors: K
authors: Nafa
authors: Department of Pathology, Memorial Sloan Kettering Cancer Center, 1275 York Avenue, New York, NY, 10065, USA. hameedm@mskcc.org.
authors: Meera
authors: M
authors: Hameed
authors: Department of Pathology, Memorial Sloan Kettering Cancer Center, 1275 York Avenue, New York, NY, 10065, USA. arcilam@mskcc.org.
authors: Marie E
authors: ME
authors: Arcila
mesh_terms: D004252:DNA Mutational Analysis / Q000379:methods* / Q000592:standards; D006801:Humans; D009154:Mutation; D015341:Nucleic Acid Probes; D009841:Oligonucleotides; D016133:Polymerase Chain Reaction / Q000379:methods* / Q000592:standards; D012680:Sensitivity and Specificity; D017422:Sequence Analysis, DNA
chemical_list: D015341:Nucleic Acid Probes; D009841:Oligonucleotides; C477371:locked nucleic acid
keywords: LNA; LNA clamping; LNA-PCR; Locked nucleic acid probes