• 来源:默瑞(上海)生物科技有限公司
  • 时间: 2020/5/12
  • 浏览人数: 1018



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

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


      “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.





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