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

Anyone used mirna mimics cholesterol conjugated for primary B cells?

I read recently that cholesterol conjugation remvoes need of lipofectamine and enhance efficiency of transfection of mimics.If any one has used please answer

11 December 2016 6,773 1 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

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

Precipitate after adding laemmli buffer to protein solution containing Potassium Acetate?

Hi, I am running a size exclusion chromatography experiment with a buffer containing Potassium Acetate as a salt. I analyse these fractions through SDS-PAGE. After boiling my SEC fractions in...

01 March 2021 2,622 3 View

What is the most suitable server for predicting protein structures in bacteria?

I have used the i-Tasser several times, however it has been unavailable for several days. I tried the swiss-model, but the output was not very pleasant due to the model used. Is there any other...

28 February 2021 4,521 3 View

Do you need the CMV enhancer before the CMV promotor for a proper function of CMV on the gene that follows?

I'm a student and I have to produce a cell line with a knockin for the NRF2 gene with GFP. I have to put a promotor in front of the GFP because the gene will be too far away from the promotor of...

28 February 2021 7,127 2 View

Are protein samples in SDS compatible with ELISA?

I have used an AllPrep DNA/RNA/Protein Mini Kit QIAGEN kit to extract RNA and protein from my samples. At the end of the protein purification, I resuspend my protein in 5% SDS. Will these samples...

28 February 2021 7,370 3 View

What is the difference between genotype and haplotype data in the ped format?

Hi everybody In the ped format for genotype, alleles of any SNP are represented by two columns (one for each allele, separated by a space). Is a column sufficient for the haplotype to...

27 February 2021 1,965 1 View

How to replicate the well conditions of a crystal hit with 20%(w/v) isopropanol?

I had a crystal hit in a well with 0.1M HEPEs pH 7.5, 10%(w/v) PEG 4000, and 20%(w/v) isopropanol. Why is isopropanol in w/v here if it's just solvent? How do I recreate this well condition?

25 February 2021 9,761 1 View

Problem with ligating 2.8 kb insert into 8.5 kb plasmid?

I am trying to clone 2 copies of the same mammalian gene into the pSF-CMV-Ub-Puro Ascl (contains HindIII, KpnI and NheI cut sites) plasmid. This is what the final product is supposed to look like...

24 February 2021 6,310 3 View

What urea buffer can i use for biotagged protein extraction from streptavidin beads?

Hi all, I have been doing research on biotagged LDB1 proteins in Mel cells. I purify the proteins using M280 streptavidin beads. The yield is very low so I've been trying to optimize it by...

24 February 2021 5,749 3 View

Can I use a protein-devoid condition as my negative control in ELISA?

Hello, I am running a sandwich-type ELISA where I am trying to inhibit a protein-protein interaction using small molecules. For each experiment, I determine my Z' (Z-prime) to determine the...

23 February 2021 7,429 1 View