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

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

Does anyone know about the inhibition of microRNA?

hello I wanna ask about inhibition of microRNA expression using vector expressing microRNA sponge. i was looking for some reference about this subject and couldn't find. can someone help me?...

10 November 2015 6,587 5 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

Why is the efficiency of viral mRNA electroporation in PBMC low?

Hello, I have been struggling with the electroporation of PBMC with HIV-I mRNA. Although the quality of the RNA generated is good, and the handling of the cells in meticulous, the frequency of...

27 February 2021 9,233 3 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

Is it efficient to do liquid biopsy to investigate the gene expressions of the tumor?

To do liquid biopsy and look for specific gene expressions there are so many published papers. Thera are different protocols and procedures used to isolate circulating cell free RNA or mRNA from...

23 February 2021 7,326 5 View

Universal bacterial primers targeting a mRNA?

Hi, I'm looking for universal primers targeting a bacterial mRNA to quatify bacterial load by qPCR in a complexe mixture (eukaryote and prokaryote RNA). Ribosomal RNA being depleted, I absolutly...

22 February 2021 9,969 3 View

How can I get the coding gene sequences of Hela and U2OS cell lines? Is mRNA sequence of Hsp90 alpha tissue specific??

I am looking for hsp90 mRNA sequence in Hela and U2OS cell lines, I have tried through ncbi but always the output is hsp90 in homo sapiens not in the specific cell line. Kindly, could any one...

22 February 2021 1,853 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

Can RNAscope be used to quantify the mRNA expression level of certain genes?

Hello, all, I want to explore the mRNA expression level of certain genes in the brain under physiological and pathological conditions. In previous studies, the researchers usually isolated the...

18 February 2021 1,518 1 View

In what ways do fellow reaerchers think that mRNA vaccination may be under CRISPR systems technology application ?

There are many ways for mRNA to intervene with genome expression. This question is raised for experts in CRISPR systems. Any RNA does not stand alone within acell. There are many ways to become...

18 February 2021 9,109 5 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