This is a guided video for RNA isolation in animal cells. If you need an explanation in Korean, you can watch it at the address below. • RNA isolation by TRIzol
Hello, thank you so much! Were any of the reagents you used cold? I've read some methodologies that use it cold Thank you
@CDRTtube
18 күн бұрын
Thank you for your question. While there may be slight variations depending on the researcher’s experience, we keep the following two reagents cold during use. Isopropanol: Used in the RNA precipitation step, and keeping it cold (around -20°C) can enhance the efficiency of RNA precipitation. Ethanol: Used in the RNA washing step, typically at a concentration of 70-75%. It is also recommended to use it in a cold state.
@MultiSupersaiyajin
17 күн бұрын
@@CDRTtube thank you!! do you think that using the trizol and chloroform in cold could harm the procedure?
@CDRTtube
17 күн бұрын
@@MultiSupersaiyajin I don't think it will have much of an effect, and since each manufacturer has a recommended temperature for storing their reagents, I think it would be a good idea to try that first.
@HRana-ej2qw
3 жыл бұрын
The reason of incomplete phase seperation after chloroform is less mixing..a good shaking of tube will mix all ingredients and forms clear 3layers
@CDRTtube
3 жыл бұрын
Thank you so much for providing valuable information. ;-) I wish you good luck with your research.
@NguyenTrung-dw2vz
2 ай бұрын
Very helpful video
@CDRTtube
2 ай бұрын
Thank you for your question. If the RNA sample is allowed to dry naturally during the final drying step, there is a risk of prolonged exposure, which could degrade the RNA. To mitigate this risk, we dry the sample quickly by applying a temperature of about 55 degrees Celsius. This method reduces RNA damage, such as RNase contamination, and does not affect the RNA quality. We have compared various drying conditions and found that this approach helps us obtain high-quality RNA. Additionally, we continuously use RNase inactivation solution to clean laboratory tables and tools. We also minimize air circulation through air conditioners to prevent RNase contamination from spreading. When necessary, we work within a biosafety cabinet. While our experience may not cover all experimental conditions, we believe you can achieve even better results by tailoring your experiments to your specific laboratory conditions. I wish you the best in all your research.
@NguyenTrung-dw2vz
2 ай бұрын
@@CDRTtube thank you so much for your detailed explain☺️
@paulineokoye8231
3 жыл бұрын
I am surprise you heated the RNA, I haven't seen or heard of such method before. isn't it going to degrade the RNA or something like that?
@CDRTtube
3 жыл бұрын
Thanks for the question. As one of the experimental methods of RNA extraction, our method uses a little heat in the last step so that the extracted RNA does not crystallize and dissolves well. In case there is a problem with the yield due to crystalization during RNA extraction, applying temperature like our method in the last step will help to obtain high-quality RNA. We wish you good results of your research. Thank you.
@bowenyang3242
3 жыл бұрын
Hi, it's a really helpful video. I am wondering about the step of after adding the chloroform. Would slightly heavier shake would be helpful to form a easier phase to extract the RNA layer?
@CDRTtube
3 жыл бұрын
Thanks for the question. After adding Trizol and chloroform, it is okay to mix vigorously. Vortexing is recommended for some experimental methods. We wish you the best results in all of your research.
@hassandastsooz3389
3 жыл бұрын
It is needed to shake strongly and vigorously. At final you will get three layers (lower phase: organic phase with pink color, interphase phase: DNA phase with white color, upper phase : aqueous phase as transparent containing RNA ).
@CDRTtube
3 жыл бұрын
@@hassandastsooz3389 Thank you for your valuable information. We wish you always have a good results. :-)
@barrierevalentin801
3 жыл бұрын
Hi, very helpful video thanks ! why do you use 75% ethanol and not 100% ethanol ?
@CDRTtube
3 жыл бұрын
Thanks for watching. When extracting RNA, precipitate it through 100% isopropanol, and add 75% Ethanol in the next step. This is to remove the salt precipitated along with the RNA during RNA extraction. So, many protocols are configured to contain between 70 and 80% ethanol. We wish you good results from all your research.
@gemijuine
3 жыл бұрын
I wish we could see how the extract looked like in an agarose gel. :D
@CDRTtube
3 жыл бұрын
Thank you for leaving a comment Next time, when RNA is extracted, a gel picture will be uploaded. We wish you will find good results for all your research.
@hadianaeemtahir8956
Жыл бұрын
this is so so helpful. Thanks a lot
@CDRTtube
Жыл бұрын
Thank you for watching. We wish you good results in all your research.
@TheMonse3000
2 жыл бұрын
Hi it's a really helpful. Thank you. How much rna dis you get?
@CDRTtube
2 жыл бұрын
Thank you for watching the video. Prices vary by country and currency exchange rate, but we usually get Trizol for around 300-350 US dollars per 100ml.
@Luxuryhairlondon
3 жыл бұрын
Hello, what’s the purpose of rnase water ?
@CDRTtube
3 жыл бұрын
Thanks for the question. Does the RNase water you asked mean RNase away? RNase away is a commercially available product. When extracting RNA, RNase away is a solution that minimizes degradation of RNA due to contamination of the solution or peripheral equipment with RNase. If you have any further questions, please feel free to ask. We hope that good research results will come from the research you are trying to do.
@HO-YTchannel
3 жыл бұрын
Woohoo! an English version of this :D
@CDRTtube
3 жыл бұрын
Thank you. Please tell us if you need an English subtitle version of another experimental video. I wish you good results.
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