How to generate single stranded florescent labelled probe for northern blotting?

Shubhangi Srivastava @Shubhangi-Srivastava

11 July 2016 1 9K Report

I want to detect small RNA, for which i need end labeled single stranded probes. what are the options for florescent labeling the probes for detection?

Vishnu Amaram

Hi Shubhangi,

There are many ways of end labeling your probes. Once you have your sequences, order the oligos and make modifications with the label you want or directly order end labeled probes from companies. Here are links for couple protocols that may be useful for you.

http://www.ncbi.nlm.nih.gov/pubmed/?term=asli+silahtaroglu%2C+sakari%2C+micrornas

http://www.ncbi.nlm.nih.gov/pubmed/?term=jove%2C+charles%2C+FISH%2C+xist

Badges
Science topic

More Shubhangi Srivastava's questions See All

Defining effective diffusivity in porous zone and non-porous zone?

Hi all, I am working on methanol steam reforming using packed bed for hydrogen generation. I am using FLUENT software for CFD simulation. In reference paper, authors mentioned that effective mass...

10 February 2021 6,104 1 View

Using two-tailed test for directional hypothesis?

In my study, I have used Process Macro to test the proposed relationships. The hypothesis is directional and two-tailed test has been deployed. Can anyone please clarify how to justify using...

06 February 2021 3,023 3 View

Which pump is used to transfer low melting temperature liquid metal from one container to another?

I want to transfer GaInSn alloy from one container to another with the help of an electromagnetic pump. Can anybody suggest me which brand or which company supplies these pumps?

25 January 2021 9,481 2 View

Can Cas13a protein directly purify from PAGE gel, please suggest me the method?

I am willing to Cas13 in vitro. Can it be directly purified from PAGE gel, please suggest me the method.

19 January 2021 1,136 3 View

I'm facing some troubling in designing a anteena ....anyone tell me how give CPW feed in cst ?

I want to give a CPW feed anyone suggest how to providea feed in antenna which I'm designed

18 January 2021 3,517 2 View

Please suggest me suitable journal of Emerald/ Elsevier / Springer for review paper.?

I am working on review paper on " Public Transportation" . Please suggest suitable journal which publishes review paper.

30 November 2020 6,687 3 View

Can any one suggest me when can i upload my paper on public forum as per the general copy right agreement ?

I published the paper one year back and received volume and issue number of my paper from publisher. Can i upload on public forum? please suggest

30 November 2020 1,219 4 View

What is your opinion about zero budget natural farming?

Agriculture is known to provide various innovations from time -to -time , but in the process , we sometimes land in no man's land, and often try to take agriculture to backfoot so miserably in...

06 November 2020 1,910 19 View

How to enforce periodic boundary condition in quantum espresso?

I want to enforce periodic boundary condition in quantum espresso. What changes do I have make in input file?

22 October 2020 6,166 2 View

JMP Pro 14 and one way ANOVA?

I have to analyse the data for one way ANOVA using JMP pro 14 software. Anyone dealt with this software or has any kind of experience. Kindly share it. Thanking you in advance.

23 September 2020 541 6 View

Gel electrophoresis result for total RNA samples, 1.5 Agarose gel was used, How to improve the integrity of RNA?

Gel electrophoresis, RNA degradation, RNA extraction from fresh tissue

02 March 2021 5,433 5 View

No title

Does anyone have the experience of using Taq Man probes in the QIAGEN Rotar- Gene qPCR machine?

01 March 2021 5,311 1 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

Any specific pipeline or parameter to analyse total transcriptomics study of brain tissue samples?

I have two groups of brain samples, control and treated for example. It was total RNA nova seq sequencing. I tried all the available pipeline like: star+rsem+deseq2, Hista+stringtie+cuffdiff,...

27 February 2021 356 6 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

Will a 10xHis-tag prevent anti-MBP from binding to MBP?

I have a protein with a 10xHis-MBP tag on the N-terminus. I'm fairly certain I am expressing my protein, but I'm not getting a signal on my western blot. Is there a possibility that the 10xHis tag...

25 February 2021 8,404 3 View

EMSA troubleshooting. Protein-DNA complex not migrating, cold probe lane with multiple bands. Can anyone help?

I'm running an EMSA to show the DNA binding activity of recombinant proteins versus the Wild type. Previous assays run by my research group using a different test system show the protein-DNA...

24 February 2021 8,325 1 View

What's the concentration recommend for recombinant protein use in western blotting?

I'm currently working having problem with the concentration for recombinant protein use in western blot, the one been used is Recombinant Human CD19 Fc Chimera Protein-rndsystem which does not...

24 February 2021 8,514 3 View

What is the best concentration of cDNA to be used for qPCR?

I have synthesised cDNA from 500ng/ul of RNA. The concentration of the cDNA Is about 5000ng/ul. Should I dilute my cDNA or just use it as it is for my qPCR? I usually use 2ul of cDNA for a 20ul...

24 February 2021 8,618 3 View

Recommendations on homogenizers for RNA isolation?

I will be performing RNA isolation with frozen lung biopsy samples and am in the market for a tissue homogenizer. I am wanting a homogenizer that will fit in a 1.5ml tube. I was looking at the...

24 February 2021 3,623 3 View