Hi, I am using TRI Reagent (Cat No TR 118), they are saying : avoid washing cells before the addition of TRI reagent as this may contribute to RNA degradation. Do you have any idea and suggestion for that?
Hi Hyeju. I'm glad I found your video! To find the best RNA concentration quality, we must do cell seeding optimization. Thus, when doing cell seeding optimization in a well plate, is it better to do it with treatment or without treatment?
i am using HEK293T cells,i washee my cells,and then trypsinized it centrifuged it and then added 1 ml trizol to the tube,and stored it on -80°c....i will perform further steps later....now someone tell me is this procedure correct or not? as i am a beginner
I add TRIzol to cells in 24- well plate to extract protein and RNA and I have 4 repeat for each treatment, but unfortunately the amount of RNA extracted from each well is very different from next well. Do you have any idea what I could do to resolve that issue?
The amount of RNA may vary depending on the number of cells seeding the cell. Next time, How about seeding on a 12well plate rather than a 24well plate?
@@hyejulee7072 but doesn’t all the unwanted salts also be precipitated if you keep the RNA in isopropanol overnight? I have heard it’s better to keep max 30 mins in ice while isopropanol precipitation of RNA.
really clear and informative vedio
It was really amazing and fun watching your videos. Thank you dear
Hi Hyeju, i really enjoyed your vid so much !!!! i got some knowledge from cell culture. thankssss
Thanks for efforts,please keep going, more videos with english subtitles.❤❤❤❤❤
Your videos are sooo cute!
Thank you sooooo much !!!
Hi, I am using TRI Reagent (Cat No TR 118), they are saying : avoid washing cells before the addition of TRI reagent as this may contribute to RNA degradation. Do you have any idea and suggestion for that?
Hi Hyeju. I'm glad I found your video!
To find the best RNA concentration quality, we must do cell seeding optimization. Thus, when doing cell seeding optimization in a well plate, is it better to do it with treatment or without treatment?
can I ask how much ng/ug of RNA do you usually get from C2C12 cells, as I saw on the eppendorf tube marking, and did you used myoblast or myotubes
i am using HEK293T cells,i washee my cells,and then trypsinized it centrifuged it and then added 1 ml trizol to the tube,and stored it on -80°c....i will perform further steps later....now someone tell me is this procedure correct or not? as i am a beginner
Great video! What kind of experiments are you doing?
Now days, I'm working on muscle atrophy using C2C12 cell, which mouse myoblast cell line !
I add TRIzol to cells in 24- well plate to extract protein and RNA and I have 4 repeat for each treatment, but unfortunately the amount of RNA extracted from each well is very different from next well. Do you have any idea what I could do to resolve that issue?
The amount of RNA may vary depending on the number of cells seeding the cell. Next time, How about seeding on a 12well plate rather than a 24well plate?
what confluency of cells is required for rna extraction?
It usually depends on the cell type and plate ! I think more than 80%.
Home made TRIzol? BTW thanks for the video.....
Our lab makes TRIzol and uses it.
isopropanol centrifugaton 60 min?
This process is necessary to centrifuge RNA pellets !
@@hyejulee7072 but doesn’t all the unwanted salts also be precipitated if you keep the RNA in isopropanol overnight? I have heard it’s better to keep max 30 mins in ice while isopropanol precipitation of RNA.