Gel Electrophoresis

Gel electrophoresis is a method for separation and analysis of macromolecules (DNA, RNA and proteins) and their fragments, based on their size and... | Contact experts in Gel Electrophoresis to get answers

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Questions related to Gel Electrophoresis

Can any one suggest me please, about a proper control for Comet Assay?

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18 March 2016 4,682 5 View

I am getting similar protein band in uninduced and induced culture. Please suggest?

The protein is an enzyme cloned in pET28a vector. I have used IPTG conc.(0.5mM, 0.8mM, 1mM) and culture at different time point (4h, 6h and overnight). I checked its induction 6 months back when...

29 February 2016 5,288 5 View

Native gel with extra Mn2+ couldnt not polymerize enough to give good wells. Why?

I was making two native gels with 5mM Mg2+ in one and Mn2+ in other. Mg2+ was fine. Mn2+ gel did not give any wells and all collapsed and remaining gel solution turned different color. Please see...

11 February 2016 3,908 1 View

Equipment and reagents required for Pulsed Field Gel Electroforesis (PFGE)?

I would like to study the molecular genetic profiling of the bacterial composition of dairy starter cultures by pulsed-field gel electrophoresis (PFGE) Any recommendation? Bio-Rad CHEF Mapper XA...

10 February 2016 4,448 4 View

Cooling module for CHEF Mapper XA just broke... Can CHEF mappers be run in 4C cold rooms?

Normally, I have 0.5 x TBE buffer running at 14C, and the cooling module keeps the buffer at this temperature. Ours seems to be broken, since it is not making the clicking noise it usually does,...

05 February 2016 319 1 View

How can you improve your 2D gel?

I am trying to run 2D gel. I am extracting protein from mammalian cell by using RIPA lysis buffer and then precipitate the protein with acetonitrile, then re suspend the protein in 250μl of...

04 February 2016 3,583 2 View

Can we perform comet assay in conventional submarine gel electrophoresis chamber?

Need a simple protocol to perform comet assay using submarine gel electrophoresis chamber (the one used for agarose gel electrophoresis).

03 February 2016 2,022 2 View

Why in my zimography, does the gel separation of animal serum show uneven staining after electrophoresis?

During the zimography after the staining gel appeared different colors separating the gel, giving us trouble in the analysis.What can I do if I am following the Krisztina Kupai protocol, PharmD,...

02 February 2016 257 9 View

Are there any suggestions for running a Native Page?

In a tube we tried the following measurements to see if it the acrylamide would polymerize before we ran the gel: 1.15mL ddH20 1mL acrylamide 0.25mL NaH2PO4 buffer 330 uL APS 33 uL TEMED Our...

30 January 2016 6,942 4 View

Has anyone ever used TGX stain-free acrylamide gels from BioRad?

Some months ago, we started using TGX Stain-Free gels (cat.# 161-0181 and 161-0185) from BioRad, and we have experienced many and different problems of polymerization. Sometimes we have to cast...

27 January 2016 3,108 3 View

I am having a problem with Tricine gels. Can no protein bands be detected at all?

I have prepared 12% Tricine gels to detect 2.2 kDa peptide band. I ran the gel but after silver staining I couldn't see any bands!!I think the gel itself is fine because I use a prestained marker...

13 January 2016 10,041 3 View

What is the alternative for Ethidium Bromide in Gel Electrophoresis?

As ethidium bromide is carcinogenic, there are several biological and environmental problems are associated with it. Is there any alternative chemical for Ethidium bromide for the detection of DNA...

08 January 2016 499 21 View

Will SDS disrupt DNA- PAGE?

Will SDS disrupt the DNA oligonucleotide PAGE? (15Bp) I have previously ran a triplex DNA, duplex DNA, single strand DNA on SDS PAGE and did not get any results. I am wondering if SDS is the...

05 January 2016 5,425 3 View

What is the minimum amount of papain (23500 Da) that I can see in the SDS PAGE gel?

I am performing reaction between iodoacetyl-PEG-biotin (linker) and papain. Thiol group of papain reacts with the linker to form a thioester bond. then I use streptavidin beads to fish out papain...

29 December 2015 4,903 3 View

Can anyone suggest a software to organize experiments from DNA isolation, PCR to Sequencing?

I need a software to organize the reactions of PCR, where all the details of the experiment are available in one software from the date the samples arrived to lab, DNA isolation, PCR, Gel...

24 December 2015 3,056 2 View

What is the share of these EtBr or SYBR in gel electrophoresis stains?

The gel electrophoresis staining market has witnessed a sharp shift from EtBr to other safer alternative products such as SYBR family of dyes, GelStar, GelRed, etc. But I think the EtBr is still...

17 December 2015 9,561 1 View

Why do my bands above the ladder? I used 100bp ladder, TBE buffer. The expected size should be 300 bp. Anyone has idea?

I am looking forward to receiving your comments and ideas and suggestions

10 December 2015 7,705 8 View

Problem with PFGE - Why is my DNA smearing?

I am attempting to work up a PFGE protocol for a Gram Positive cocci that is of interest to our lab. However, I am still getting a lot of undigested material stuck up in the loading well. I...

25 November 2015 7,067 7 View

Should I store the Tricine gels in an annode buffer or can I use the running buffer from the Laemmli system?

I'm wondering how other people store their handcast Tricine gels. The Laemmli gels we make are usually stored in a container of running buffer. Should I store the Tricine gels in annode buffer? Or...

20 November 2015 3,902 2 View

Any advice on denaturing urea PAGE gel electrophoresis staining?

Dear All, After a successful amplification PCR I am about to visualize my AFLP fragments. Most of the protocols suggests UREA PAGE. The protocol i used ...

22 October 2015 6,835 4 View

Is it ok to select all lanes in one rectangle and draw vertical line to seperate each lane for gel image analysis using ImageJ?

Hello, For using ImageJ to analyse gel image, normally I select each lane for each rectangle/box. but some guides suggest to draw a rectangle around all the lanes of gel image in one box and then...

07 October 2015 8,127 5 View

How could I check whether there is RNA in my protein solution?

I purified a recombinant RNA binding protein from E.coli, I worried that my RNA-binding protein was contaminated with host RNA during purification. I need to remove RNA completely, I have treated...

07 September 2015 9,464 12 View

How can I amplify a low concentration DNA target within a high DNA concentration solution?

I am currently trying to detect bacterial 16S rRNA DNA in human tissue to ultimately sequence the results. I conducted PCR on diluted samples (original concentration of DNA was between...

26 August 2015 341 5 View

How can I eliminate non-specific amplification during SYBR qPCR?

I have been designing qPCR primers for use in assays with Promega GoTaq qPCR mastermix. This uses BRYT Green, which is supposed to be comparable to SYBR chemistry. Using a non-precious positive...

26 August 2015 1,624 8 View

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