which vector is best to express a pre-miRNA? Can I buy it in addgen?
Thanks for your suggestion , I will try. I am now trying a regular vector- pEGFP-c1, I hope it can work.
I want to purify a 6XHis-tagged protein from 293T cells, what lysis buffer should I use? After the cell lysis I will use Nickel Resin to purify this protein. Thanks
31 December 2014 9,535 4 View
We have constructed several plasmids that express HA- tagged proteins. At first, all the protein expression of these plasmids were confirmed by WB. But now, after several rounds of amplification...
09 October 2012 6,837 6 View
We want to buy a DNA/RNA synthesizer, which can produce DNA primer, single strand and double strand RNA oligos with various modifications. I don't know which model is easy to use and reliable, are...
09 October 2012 6,530 5 View
Taqman or LNA, or any other methods? If I chose one of these methods, should I also buy their related products, like RT kit and Real-time PCR assay kit, too?
02 March 2012 6,890 10 View
Dear All, mirna primer showing some problem in the melting curve? any idea why? As attached is the melting curve. The forward sequence is obtained from miRBase and reverse primer is universal.
28 February 2021 5,008 4 View
In diseased conditions, many genes and gene products are visible in the bloodstream. So my question is whether we can find traces of microRNA which is involved in disease and disorder in blood serum?
24 February 2021 7,064 3 View
Hello RG community, I wonderer whether exogenous molecules (e.g. reporter plasmids, miRNAs or other artificial molecules) are transferred between cultured human cells during their doubling ? So,...
21 February 2021 6,633 3 View
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...
17 February 2021 8,986 2 View
I need to quantify blood plasma derived miRNA through q-RT-PCR. For this I need to fix RNA content in my cDNA reaction. What is the best way to do this? Currently, I take readings of my plasma...
03 February 2021 9,330 3 View
I am using the Qiagen miRNEASY serum/plasma advanced kit, using 400ul of frozen plasma. What is the yield you get on average? I am using nanodrop currently for quantification. I am getting...
02 February 2021 8,806 2 View
I'm working on selecting mouse miRNAs to analyze and stumbling upon one confusing issue, some of the papers do not specify which arm they analyzed and they just state something like " miR-34a was...
16 January 2021 3,792 4 View
I am trying to confirm my miRNA target by western blotting. I have a ds mimic and a ss inhibitor of my miRNA. Is it necessary to use negative controls for mimic and inhibitor? If yes, how can I...
09 January 2021 2,320 3 View
Hello! I am trying to predict in silico the miRNA by knowing the target. Most of published research starts by knowing the miRNA and predicts the target using online tools. Are you familiar with...
31 December 2020 7,433 5 View
I am searching for miRNA repeats put between WPRE and 3'LTR of my lentivirus, in order to enhance the specificity of transgene expression in human neuron cells. But if clinically translation of...
26 December 2020 9,921 2 View