How mirna chose its target among 100s of mRNA having seed sequence?

I am not clear what is rule behind mirna and mRNA targeting pair,because each mirna target officially 100s of target. In such cases how one will validate legitimate mirna mRNA target. Assays...

03 April 2019 7,825 5 View

How to meassure primary miRNA trancript using QRT PCR?

I want to know how to design primer for primary miRNA QRT PCR

02 March 2018 220 7 View

Can I store phenol chloroform phase in trizol?

I use trizol for RNA isolation,but we store phenol choroform phase -30 , I want to know is it good to do isolation of protein later after storage -30 of the above mentioned phase or it may affect...

10 November 2017 7,410 0 View

Can miRNA mimics or inhibitor used in primary cells?

I am currently working with lymphocyte ,I want to know if any one has used the miRNA mimics or inhibitor with primery cells

10 November 2016 3,344 4 View

Can i use different concentration of forward and reverse primer in real time ?

i am currently making assay of syber green based real time pcr for detecting rare isoforms.To make them specific ,can i use say 1 p mole of forward and 5 p mole of reverse primer,technically for...

06 July 2016 8,260 4 View

Can anyone provide a table for miRNA and respective gene from which it arises?

There are certain miRNA originated from the intron of genes. I could not found such tabulated form in which genes and respective miRNA are given Can anyone provide the same.

11 December 2015 1,564 3 View

In rel time PCR non specific band at higher dilution?

09 October 2015 6,164 3 View

What is meaning of miR17 approximately 92 cluster?

During literature search I found miR 17/92,miR-17 approximately 92, miR 17~92 I am not able to understand practical meaning of these annotations in microRNA. Can anybody help to me to understand...

09 October 2015 6,743 1 View

Can anyone help me with the interpretation of intron centric analysis file output ?

RNA seq data analysis by intron centric analysis gives insight about exon skkiping ,I have attached photo of excel file I have. Can anyone help me how to interpretate it eg...

06 July 2015 9,479 4 View

What are different algorithms to analyse microarray data?

I need to analyse microarray data ,so I need latest algorithms can any body help me to find ..

05 June 2015 7,282 0 View

Why might my GFP tag be dissociating from my plasmid after transfection?

I have a set of stably transfected cell lines all transfected with plasmids containing GFP tags on the C terminus. During a western blot using anti-GFP antibody, one of my plasmids has dissociated...

01 March 2021 9,310 4 View

Can someone suggest a way to prepare slides to observe under confocal microscope for observing fluorescent tagged proteins??

I am growing the cells on coverslips and transfecting the fluorescent tagged protein directly on them. Then i want to observe these cells under confocal microscope. Currently i am just using...

01 March 2021 6,142 3 View

Problem with shRNA transfection. Why are only two plates getting contaminated instead of all?

I transfected my LNCaP-WT cells with 3 shRNA plus their NTC two weeks ago and split two puromycin selected cell plates on Friday last week(Feb 26). I checked for GFP in the cells, and they all...

28 February 2021 4,949 3 View

Does RNAiMAX transfection reagent work in serum free medium?

I diluted siRNA and RNAiMAX in opti-MEM and added to the cells which they were in the growth medium. Is it a right way? or should I culture cells in the opti-MEM medium for a while and not in...

26 February 2021 10,041 3 View

How do I transfect a large plasmid into PC12 cells with nerve growth factor (NGF) receptor?

I have a large 16 kb plasmid, which I need to transfect into PC12 cells. Lipofectamin 2000 didn't work and with GFP alone the transfection rate is very low. I also tried the neon invitrogen...

25 February 2021 6,635 5 View

What is a good reagent for lentiviral plasmid transfection in HeLa cells?

I have been using Lipofectamine 2000 but was hoping there may be a better protocol or one for something like Lipofectamine PLUS reagent.

24 February 2021 1,633 4 View

Why the cells die after stable transfection?

I tried stable transfection in the U87 cell line with the sgRNA-Cas9 All-in-One lentivector. Cells were cultured in a 6-well plate, 4ug plasmid was transfected after 24hs when the cell confluence...

22 February 2021 2,864 4 View

Is there a method for evaluating nanoparticle adhesion to epithelial tissue in vitro?

I want to measure how well nanoparticles can transfect cells based on NP adhesion properties. In the literature I find people do it in animal studies, and epithelial adhesion helps in transfection...

21 February 2021 7,791 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

Which transfection reagent is the best for plasmid transfection of MDA-MB-231 cell line, Lipofectamine based or other reagent suggestion?

I try to transfect MDA MB 231 cell line with a plasmid by PEI, unfortunately, the rate of transfection is low, and now I wanna find and order the best transfection reagent for this cell line and...

19 February 2021 828 6 View