TWISTDX RPA試劑盒（（加MMLV和抑制劑）操作建議

英國TwistDx公司（前身為ASM科技有限公司）成立于1999年，基于英國劍橋Babraham研究院建立的生物技術(shù)公司。該公司通過突破等溫DNA擴(kuò)增，開發(fā)的重組酶聚合酶擴(kuò)增*技術(shù)（RPA），被譽(yù)為DNA診斷領(lǐng)域的革命性創(chuàng)新。

RT試劑盒舉例

（1）Revert Reverse Transcriptase(200U/uL)(Thermo EP0441)

（2）RNA Inhibitor (40,000 U/mL)( Enzymatics Y9240L

TWISTDX廠家回復(fù)

　　“We recommend using similar RT amounts to that of a PCR reaction of the same volume, according to your chosen manufacturer’s instructions.

　　We also suggest that you run reactions at 40°C and delay agitation of the reaction by a minute to give the RT time to work, otherwise you just run the reaction as you would a normal TwistAmp® reaction (it’s a one-step process).

　　It’s also advisable to add RNase Inhibitor to any reaction where RNA is the target material. Any commercially available RNase Inhibitor will be suitable.

　　If the M-MLV RT that you are using comes with a separate dilution/reaction buffer, we suggest not using it as it will lower the concentration of the enzyme.

　　The “delay agitation of the reaction by a minute” refers to the first mixing step per the protocol in the manual (Step 4). So wait 5 minutes instead of 4 minutes before “vortex, spin down briefly, and return the samples to the reader ensuring that the tubes are returned to their original positions in the block.”

　　If we were to try and use it again, we would do a titration to determine optimal quantity to add in the new formulation reactions (we haven’t used it since we changed the formulation of the reactions a couple of years ago).

　　When using RNase Inhibitor. We have used 1µg /50µl reaction, some R&D users have used 2µg /50ul, which didn’t appear to have a negative effect on the performance in our own assay.

　　For this : RNAse Inhibitor, that manufacture uses 10 ul of the RNAse solution in 50 ul reaction volumes for activity determination, but your customer can try different amounts 5ul or 20ul to see what will give the best results. For this product, references of 50 ul PCR reaction volumes would also be reasonable.

TwistDx以此技術(shù)為基礎(chǔ)生產(chǎn)的核酸擴(kuò)增類產(chǎn)品，能在恒定且較低的溫度下，僅用10~20分鐘，進(jìn)行單分子核酸檢測(cè)。這一技術(shù)對(duì)環(huán)境、硬件設(shè)施的要求很低，對(duì)于生物防護(hù)、水體檢驗(yàn)、食品檢驗(yàn)、醫(yī)療診斷、微流體、獸醫(yī)等方面都有良好的應(yīng)用。

RPA擴(kuò)增的各種體系試劑盒中，含有DNA、RNA擴(kuò)增所需的所有試劑，研究者只要準(zhǔn)備好引物和模板就行了。擴(kuò)增結(jié)果可以分別通過凝膠電泳檢測(cè)、熒光定量檢測(cè)或試紙條檢測(cè)，產(chǎn)物純化后可用于下游研究。

默瑞生物

默瑞(上海)生物科技有限公司，作為專業(yè)的生命科學(xué)產(chǎn)品推廣與服務(wù)商，以“創(chuàng)新，專業(yè)，共贏”的理念，不斷強(qiáng)化服務(wù)體系，提升客戶體驗(yàn)，以莫納生物產(chǎn)品為主線，結(jié)合國內(nèi)外優(yōu)質(zhì)產(chǎn)品提供高標(biāo)準(zhǔn)完整解決方案，幫助科研客戶和生物技術(shù)企業(yè)高效創(chuàng)新，加速轉(zhuǎn)化，創(chuàng)造更多優(yōu)質(zhì)產(chǎn)品，提升社會(huì)福祉，實(shí)現(xiàn)多方共贏。

　　默瑞生物創(chuàng)立以來，已經(jīng)陸續(xù)導(dǎo)入TwistDx,enzymatics,KAPABIOSYSTEMS,BioDynami,Jackson,Echelon,ChromoTek,Nanocs,Biotechrabbit,LeeBiosolutions, magtivio等優(yōu)質(zhì)品牌，逐步培育成為專業(yè)用戶的選擇。