I've been trying to quantitate mature microRNAs in 293T cells with and without 'X'. 2ug of total RNA (isolated by Trizol method) is converted to cDNA using All-in one microRNA cDNA synthesis kit (abm good) and the abm primers are used to quantify the expression of mature miRs. Has anyone had any problems with the company's products ? Im not usuing the traditional kit, so some reviews on the products would be great!
Thanks much!
hi,
Are you looking for a product review or encountered a problem while or after using?
I have followed a similar protocol/steps but used the Qiagen miScript RT II kit. Well, the Qiagen kit is compatible with the SYBr master mix. I found the Thermo miRNA advanced cDNA kit works with Taqman. So, here, in your case check the master mix compatibility.
Use good 2-3 endogenous control for normalization.
Check, papers for default expression, most important what is the miRNA level in HEK293 cells.
Regards
Saurabh
Thanks for the prompt response, Saurabh Mandal
I was looking for a product review. I haven't encountered problems as such with the abm good kits. It's just that I came across a couple of comments on research gate stating their inefficiency ( primer-dimers and false results!) which got me concerned.
I am currently using RNU43 as my control. Will add U6 and SNORD44 to the panel.
I will also look into their basal level expressions as suggested.
Thanks for your valuable comment ! I appreciate it.
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