Q&A

  • Q:

    Column blockage

    A-1 Cell lysis or homogenization not sufficient

    ---- Reduce sample usage, increase the amount of lysis buffer, increase homogenization and lysis time.


    A-2 Sample amount is too large

    ---- Reduce the amount of sample used or increase the amount of lysis buffer.

  • Q:

    Low RNA yield

    A-1  Insufficient cell lysis or homogenization

    ---- Reduce sample usage, increase the amount of lysis buffer, increase homogenization and lysis time.


    A-2  Sample amount is too large

    ----Please refer to the maximum processing capacity.


    A-3 RNA is not eluted completely from the column

    ---- After adding RNase-Free water, leave it for a few minutes before centrifuging.


    A-4 Ethanol in the eluent

    ---- After rinsing, centrifuge again and remove the washing buffer as much as possible.


    A-5 Cell culture medium is not completely removed

    ---- When collecting cells, please make sure to remove the culture medium as much as possible.


    A-6 The cells stored in RNAstore are not effectively centrifuged

    ----RNAstore density is greater than the average cell culture medium; so the centrifugal force should be increased. It is suggested to centrifuge at 3000x g.


    A-7 Low RNA content and abundance in the sample

    ---- Use a positive sample to determine if the low-yield is caused by the sample.

  • Q:

    RNA degradation

    A-1  The material is not fresh

    ---- Fresh tissues should be stored in liquid nitrogen immediately or immediately put into the RNAstore reagent to ensure the extraction effect.


    A-2  Sample amount is too large

    ---- Reduce sample amount.


    A-3  RNase contamination

    ----Although the buffer provided in the kit does not contain RNase, it is easy to contaminate RNase during extraction process and should be handled with care.


    A-4  Electrophoresis pollution

    ---- Replace the electrophoresis buffer and make sure the consumables and Loading Buffer are free of RNase contamination.


    A-5  Too much loading for electrophoresis

    ---- Reduce the amount of sample loading, the loading of each well should not exceed 2 μg.

  • Q:

    DNA contamination

    A-1  Sample amount is too large

    ---- Reduce sample amount.


    A-2  Some samples have high DNA content and can be treated with DNase.

    ----Perform RNase-Free DNase treatment to the obtained RNA solution, and the RNA can be directly used for subsequent experiments after treatment, or can be further purified by RNA purification kits.

  • Q:

    How to remove RNase from experimental consumables and glasswares?

    For glasswares, baked at 150°C for 4 h. For plastic containers, immersed in 0.5 M NaOH for 10 min, then thoroughly rinsed with RNase-free water and then sterilize to completely remove RNase. The reagents or solutions used in the experiment, especially water, must be free of RNase. Use RNase-free water for all reagent preparations (add water to a clean glass bottle, add DEPC to a final concentration of 0.1% (V/V), shake overnight and autoclave).