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Cfx96 real time pcr

Manufactured by Thermo Fisher Scientific
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The CFX96 Real-Time PCR is a laboratory instrument used for quantitative polymerase chain reaction (qPCR) analysis. It is capable of performing real-time PCR experiments to accurately measure and quantify target DNA or RNA sequences.

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Lab products found in correlation

Mutation detector software, by Thermo Fisher Scientific (2 mentions) Maxwell dna ffpe kit, by Promega (2 mentions) Taqman mutation detection assay, by Thermo Fisher Scientific (2 mentions)

2 protocols using cfx96 real time pcr

1

Validating Low Frequency Variants using TaqMan Assays

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Low frequency variants for which commercial primers were available were validated with TaqMan Mutation Detection Assays (Thermo Fisher) (supplementary figure S1). Mutation detection was performed according to assay instructions. Briefly, FFPE tissue was micro-dissected from multiple regions of each tumor (supplementary figure S1A-C) under a dissection microscope and DNA was purified using the Maxwell DNA FFPE Kit (Promega). Samples were run in duplicate or triplicate per manufacturer’s instructions to determine the presence of or the frequency of the variant DNA, respectively. The qPCR reactions were run on Bio-Rad CFX96 Real-Time PCR and data was analyzed using the Mutation Detector™ software (Thermo Fisher, last revised April 2012). Variants that fell below our variant calling cutoffs for sequencing, but that were validated by qPCR were included in the data set. This included the KRAS variant for polyp PF24 and the removal of the CTNNB1 variant in PF11.
Sievers C.K., Zou L.S., Pickhardt P.J., Matkowskyj K.A., Albrecht D.M., Clipson L., Bacher J.W., Pooler B.D., Moawad F.J., Cash B.D., Reichelderfer M., Vo T.N., Newton M.A., Larget B.R, & Halberg R.B. (2016). Subclonal diversity arises early even in small colorectal tumours and contributes to differential growth fates. Gut, 66(12), 2132-2140.
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2

Validation of Low-frequency Genetic Variants

Check if the same lab product or an alternative is used in the 5 most similar protocols
Low-frequency variants for which commercial primers were available were validated with TaqMan Mutation Detection Assays (Thermo Fisher) (see online supplementary figure S1). Mutation detection was performed according to assay instructions. Briefly, FFPE tissue was microdissected from multiple regions of each tumour (see online supplementary figure S1A–C) under a dissection microscope and DNA was purified using the Maxwell DNA FFPE Kit (Promega). Samples were run in duplicate or triplicate as per manufacturer's instructions to determine the presence of or the frequency of the variant DNA, respectively. The qPCR reactions were run on Bio-Rad CFX96 Real-Time PCR and data were analysed using the Mutation Detector software (Thermo Fisher, last revised April 2012). Variants that fell below our variant calling cut-offs for sequencing, but that were validated by qPCR were included in the dataset. This included the KRAS variant for polyp PF24 and the removal of the CTNNB1 variant in PF11.
Sievers C.K., Zou L.S., Pickhardt P.J., Matkowskyj K.A., Albrecht D.M., Clipson L., Bacher J.W., Pooler B.D., Moawad F.J., Cash B.D., Reichelderfer M., Vo T.N., Newton M.A., Larget B.R, & Halberg R.B. (2016). Subclonal diversity arises early even in small colorectal tumours and contributes to differential growth fates. Gut, 66(12), 2132-2140.
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