Anchored PCR (A-PCR) for genome walking?

i am trying to identifies unknown sequence in nitrate reductase gene from chlorella vulgaris using anchored pcr. first i extracted the genomic DNA and i did single primer extension using 0.5 ug of...

05 June 2018 8,739 11 View

DNA and RNA extraction from chlorella vulgaris?

hello i am trying to extract DNA and RNA from chlorella vulgaris. first i tried the CTAB methods but i couldn't get high yields and the quality of DNA or RNA. I used the TaKaRa MiniBEST Plant...

03 April 2018 6,663 10 View

Overlapping pcr of 4 fragments?

hello i am trying to overlap four fragment through use of overlapping pcr: the fragments are CMV promoter (595bp), Kanamycine resistance gene (795bp), IRES (206), and EGFP (720bp). i designed the...

01 February 2017 9,201 5 View

Live modified organism?

hello i was reading this paper http://www.cell.com/trends/biotechnology/abstract/S0167-7799(15)00088-8 and they used the cas9 protein and guide RNA without use of plasmid. is it possible to call...

10 November 2016 661 13 View

Protein Electrophoresis and Blotting for RNA page electrophoresis?

06 July 2016 1,983 3 View

What happen to microrna after inhibiting translation of mRNA?

hello everyone i am curious about the fate of microrna after inhibiting the translation of mRNA can someone provide me with information about it thanks in advance.

03 April 2016 471 0 View

Extract 3'utr from annotation file?

hello i am looking to extract the 3'utr sequences of fathead minnow fish using the annotation file and the genome. http://www.ncbi.nlm.nih.gov/genome/?term=fathead%20minnow can someone help me or...

02 March 2016 6,470 0 View

Construction of artificial microrna vector to target a gene?

31 December 2015 2,661 4 View

MicroRNA blast against miRbase?

Hello everyone I did an experiment using microRNA NGS and because of lack of funds we did the alignment of microRNA against zebarfish microrna and I got some novel microRNA. Now I want to...

11 December 2015 9,701 6 View

For MiRNA knockout, should I use a bulged microRNA sponge or a perfectly complementary one?

hello everyone i wanna knockout a specific microRNA and i would use microRNA sponge and through my readings i found some researchers used bulged sponge and others used perfect sponge complementary...

10 November 2015 5,602 0 View

MiRNA qpcr problem?

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

Can MicroRNA cross the blood brain barrier?

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

Are exogenous molecules transferred between human cells during doubling ?

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

Has anyone encountered any problems with abm good microRNA primers for quantification of mature miRNAs?

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

What is the best way to fix plasma derived miRNA for subsequent cDNA synthesis?

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

What is the general miRNA yield one gets from the Qiagen miRNEASY serum/plasma advanced kit?

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

Which miRNA arm is a right target for analysis, 3p or 5p?

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

Is negative controls necessary for western blotting?

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

Can I predict miRNA by knowing the mRNA target?

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

What are the possible drawbacks of using miRNA sponge to enhance the specificity of gene expression?

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