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Therascreen kras mutation kit

Manufactured by Qiagen
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TheraScreen KRAS mutation kit is a lab equipment product designed for the detection of KRAS gene mutations. It is used for the analysis of KRAS mutation status in DNA samples.

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Lab products found in correlation

Rotor gene q, by Qiagen (1 mentions) Ffpe dna isolation kit, by Qiagen (1 mentions) Rotor gene q real time pcr platform, by Qiagen (1 mentions)

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TheraScreen DxS KRAS Mutation Kits KR-21 and KR-22 (2 citations)

4 protocols using therascreen kras mutation kit

1

KRAS Mutation Detection via Scorpion-ARMS PCR

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Scorpion-ARMS real-time PCR (TheraScreen KRAS mutation kit, Qiagen) was used according to the manufacturer’s instructions. This assay is designed to detect a wild-type control and seven KRAS exon 2 mutations: G12A, G12D, G12R, G12C, G12S, G12V, and G13D. Real-time PCR was performed using the Rotor-Gene Q instrument (Qiagen). Data regarding each mutation was interpreted according to the kit manual after a curve analysis and the calculation of the δCt values (sample mutation assay Ct minus sample control assay Ct). The manufacturer has reported the sensitivity to be 1% mutant alleles in a wild-type background if a sufficient DNA input is used.
Sakai K., Tsurutani J., Yamanaka T., Yoneshige A., Ito A., Togashi Y., De Velasco M.A., Terashima M., Fujita Y., Tomida S., Tamura T., Nakagawa K, & Nishio K. (2015). Extended RAS and BRAF Mutation Analysis Using Next-Generation Sequencing. PLoS ONE, 10(5), e0121891.
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2

KRAS Mutation Detection in FFPE Samples

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DNA was isolated from 10-μm formalin-fixed, paraffin-embedded tumor specimens using FFPE-DNA isolation kit (Qiagen, Hilden, Germany). A Qiagen the rascreen KRAS mutation kit was used to detect the seven most common KRAS codon 12 and 13 mutations. Specifically, the mutation was detected by real-time polymerase chain reaction based on amplification-refractory mutation system and Scorpion probes (Gly12Asp [GGT > GAT] G12D, Gly12Val [GGT > GAC] G12V, Gly12Cys [GGT > TGT] G12C, Gly12Ser [GGT > AGT] G12S, Gly12Ala [GGT > GCT] G12A, Gly12Arg [GGT > CGT] G12R, Gly13Asp [GGC > GAC] G13D).
Kim H.S., Heo J.S., Lee J., Lee J.Y., Lee M.Y., Lim S.H., Lee W.Y., Kim S.H., Park Y.A., Cho Y.B., Yun S.H., Kim S.T., Park J.O., Lim H.Y., Choi Y.S., Kwon W.I., Kim H.C, & Park Y.S. (2016). The impact of KRAS mutations on prognosis in surgically resected colorectal cancer patients with liver and lung metastases: a retrospective analysis. BMC Cancer, 16, 120.
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3

KRAS Mutation Detection Protocol

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A Therascreen KRAS Mutation Kit (Qiagen) was used according to the manufacturer's instructions. This assay is designed to detect a wild-type control and the seven most common KRAS mutations in codons 12 and 13 of exon 2, as shown in table 1. Real-time PCR was performed on a Rotor-Gene Q Real-time PCR Platform (Qiagen). The cycling conditions for quality control runs and mutation assays were as follows: 15 min at 95°C, followed by 40 cycles at 95°C for 30 s and 60°C for 1 min. Fluorescence was measured at 60°C. Data on each mutation were interpreted according to the kit manual after curve analysis and calculation of ΔCt values.
Gao J., Wu H., Wang L., Zhang H., Duan H., Lu J, & Liang Z. (2016). Validation of targeted next-generation sequencing for RAS mutation detection in FFPE colorectal cancer tissues: comparison with Sanger sequencing and ARMS-Scorpion real-time PCR. BMJ Open, 6(1), e009532.
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4

Detection of KRAS Mutations in Tumor Samples

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The TheraScreen K-RAS Mutation Kit (Qiagen N.V., Venlo, Netherlands) was used to detect somatic mutations in KRAS genes. One paraffin block containing a high density of tumor cells was selected from each case, from which H-E-stained slides were reviewed by a pathologist (M.T.). For each case, 10 unstained, 10-μm thick sections were cut and mounted on aminopropyltriethoxysilane-coated slides. The region with the highest density of tumor cells was selected for microdissection and its targets were confirmed by direct microscopic visualization. Tumor DNA was extracted from the microdissected fragments. Real-time polymerase chain reaction was performed to analyze seven mutations (Gly12Ala, Gly12Asp, Gly12Arg, Gly12Cys, Gly12Ser, Gly12Val, and Gly13Asp). An amplification refractory mutation system was used for allele-specific amplification, which was detected using Scorpion primers. Based on gene analysis, tumors were classified into KRAS wild-type (KRAS WT ) or KRAS mutant-type (KRAS MT ).
Sakanashi F., Shintani M., Tsuneyoshi M., Ohsaki H, & Kamoshida S. (2019). Apoptosis, necroptosis and autophagy in colorectal cancer: Associations with tumor aggressiveness and p53 status. Pathology, research and practice, 215(7).
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