Real-time Polymerase Chain Reaction:Article Title: High-throughput informative single nucleotide polymorphism-based typing of Neisseria gonorrhoeae using the Sequenom MassARRAY iPLEX platform.
Article Snippet: .. In this study, an iPLEX MassARRAY method (iPLEX14SNP) was developed targeting 14 informative SNPs across six (abcZ, aroE, fumC, gdh, pdhC and pgm) of the seven gonococcal housekeeping genes that are commonly used for MLST investigations.5 In brief, the iPLEX14SNP assay was an adaption to the Sequenom platform of a previous real-time PCR 14 SNP method described by our laboratory, which we showed could be used to individually distinguish 155 of 217 MLST types in the MLST database (database accessed on 20 June 2012), with a further 62 MLST types having overlapping 14 SNP profiles (between two and four MLST types with the same 14 SNP profile).5 Amplification primers and extension primers (Tables 1 and 2) for each of the 14 SNPs in the iPLEX14SNP were N. gonorrhoeae SNP-based typing JAC D ow nloaded from https://academ ic.oup.com /jac/article/69/6/1526/830926 by guest on 16 June 2024 designed using the Assay Designer 4.0 software (Sequenom) and based on methods previously described.10 All amplification primers commenced with the 5′ 10-mer tag (ACGTTGGATG) to ensure they fell outside the observed mass window. .. The initial amplification PCR was performed on a conventional 96-well thermal cycler (Applied Biosystems, USA) using the Sequenom amplification kit.
Amplification:Article Title: High-throughput informative single nucleotide polymorphism-based typing of Neisseria gonorrhoeae using the Sequenom MassARRAY iPLEX platform.
Article Snippet: .. In this study, an iPLEX MassARRAY method (iPLEX14SNP) was developed targeting 14 informative SNPs across six (abcZ, aroE, fumC, gdh, pdhC and pgm) of the seven gonococcal housekeeping genes that are commonly used for MLST investigations.5 In brief, the iPLEX14SNP assay was an adaption to the Sequenom platform of a previous real-time PCR 14 SNP method described by our laboratory, which we showed could be used to individually distinguish 155 of 217 MLST types in the MLST database (database accessed on 20 June 2012), with a further 62 MLST types having overlapping 14 SNP profiles (between two and four MLST types with the same 14 SNP profile).5 Amplification primers and extension primers (Tables 1 and 2) for each of the 14 SNPs in the iPLEX14SNP were N. gonorrhoeae SNP-based typing JAC D ow nloaded from https://academ ic.oup.com /jac/article/69/6/1526/830926 by guest on 16 June 2024 designed using the Assay Designer 4.0 software (Sequenom) and based on methods previously described.10 All amplification primers commenced with the 5′ 10-mer tag (ACGTTGGATG) to ensure they fell outside the observed mass window. .. The initial amplification PCR was performed on a conventional 96-well thermal cycler (Applied Biosystems, USA) using the Sequenom amplification kit.
Software:Article Title: High-throughput informative single nucleotide polymorphism-based typing of Neisseria gonorrhoeae using the Sequenom MassARRAY iPLEX platform.
Article Snippet: .. In this study, an iPLEX MassARRAY method (iPLEX14SNP) was developed targeting 14 informative SNPs across six (abcZ, aroE, fumC, gdh, pdhC and pgm) of the seven gonococcal housekeeping genes that are commonly used for MLST investigations.5 In brief, the iPLEX14SNP assay was an adaption to the Sequenom platform of a previous real-time PCR 14 SNP method described by our laboratory, which we showed could be used to individually distinguish 155 of 217 MLST types in the MLST database (database accessed on 20 June 2012), with a further 62 MLST types having overlapping 14 SNP profiles (between two and four MLST types with the same 14 SNP profile).5 Amplification primers and extension primers (Tables 1 and 2) for each of the 14 SNPs in the iPLEX14SNP were N. gonorrhoeae SNP-based typing JAC D ow nloaded from https://academ ic.oup.com /jac/article/69/6/1526/830926 by guest on 16 June 2024 designed using the Assay Designer 4.0 software (Sequenom) and based on methods previously described.10 All amplification primers commenced with the 5′ 10-mer tag (ACGTTGGATG) to ensure they fell outside the observed mass window. .. The initial amplification PCR was performed on a conventional 96-well thermal cycler (Applied Biosystems, USA) using the Sequenom amplification kit.
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