EGFR突變檢測試劑盒 | Detection kit for EGFR Mutations, BD0040
EGFR突變檢測試劑盒 (Detection kit for EGFR Mutations, BD0040)(螢光探測)採用螢光探查技術,檢測肺癌患者EGFR六種突變類型;適用於血清(serum)、血漿(plasma)或非小細胞組織(non small cell lung cancer )。檢測結果可以反映易瑞沙(iressa)和特羅凱(tarceva)等靶向藥物的有效性,但不用於臨床診斷(clinical diagnosis)和治療。【檢測原理】 該試劑盒使用 TaqMan-MGB 探針技術(TaqMan-MGB probing technology)檢測六個突變位點。 5'TaqMan-MGB 探針的末端signed by reporter such as FAM, VIC。 MGB 可以增強探針與模板,提高探針的 Tm。這種帶有 MGB 的較短探頭可以達到更高的Tm 類似,使報告基因更接近猝滅劑,從而產生更好的猝滅效果,更低的螢光背景和更高的信噪比(signal-noise ratio)。探測完成當它不與模板結合時,另外,由報告基因被淬滅劑吸收,導致不發螢光。否則,探針在與模板結合時被 Taq 的 5'→3' 切除酶降解,導致報告分子和猝滅劑分離,然後發出螢光出現了。 TaqMan-MGB 探測方法具有一些優點,例如更高的敏感性和特異性。一管多重PCR,可減少生產成本和提高效率,可以通過替代實現不同波長的螢光組。
Procedure :
[Self-contained reagents] Tips, PCR 96 pore plates or eight reaction tubes without RNase and DNase. [Suitable instrument] ABI PRISM 7000/7700/7300, StepOnePlus Real-Time PCR System and so on. [Requirement samples] Detectable samples are as follows: 1. Serum, blood, plasma DNA. 2. Tissue including fresh or embedded in paraffin, operation sample conserved in liquid nitrogen, ethyl alcohol and RNAlater. 3. Sample from puncture and biopsy. DNA extraction: Using commercial kit to extract human genomic DNA such as QIAamp DNA Blood Mini Kit. Quantifying extracted DNA and adding less than100ng template in 20μL system. [Calibration method] In every PCR reaction, sample, positive and negative control need analyze simultaneously. 1. Preparing reagent for reaction Taking out nucleic acid reaction solution from the kit and thawing, then centrifuging at 2000rpm for 10sec. 2. Loading samples a). Adding 7.6μL PCR reaction, 10μL 2×Taq Mix and 0.4μL ROX Reference Dye to every tube. Reaction liquid can also be mixed and packaged subsequently. b). Adding 2μL DNA template, and quantifying DNA under 100ng firstly. c). Centrifuging all reagents in PCR 96 pore plates or eight reaction tubes to the bottom. 3. PCR amplification a). Putting PCR reaction plates in fluorescence quantitative PCR instrument. b). Cyclic condition setting
For research use only, not for use in diagnostic procedures. The validity of reagents in kit is six months (Please use within the validity period ).
Bioworld台灣代理商-BRAND/AGENCY/代理品牌/ Bioworld台灣代理商
Bioworld擁有專業經驗豐富的科學和技術專業人員,製造和銷售抗體。 我們目前的抗體覆蓋範圍廣泛的信號轉導和細胞挑選,例如:
1)染色質和酰化信號
2)MAPK信號
3)神經元SAPK / JNK信號
4)細胞凋亡和Caspase信號
5)AKT信號
6)轉錄和翻譯控制
7)PKC信號
8)細胞週期和DNA損傷信號
9)細胞骨架信號