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

Does the E.coli DH5 alpha have an inherent ligase?

Does the E.coli DH5 alpha have an inherent ligase? If yes, is it possible that a restricted digested plasmid would get recircularized using the ligase from the E.coli DH5 alpha once its...

07 July 2014 9,958 3 View

Why do I get the cloned insert size rather than the plasmid size when I gel analyze after maxiprep?

I did maxiprep plasmid/BAC isolation. When I did gel analyze, I was expecting for the classical two bands for the supercoiled and linear plasmids. Instead, I got real intense bands that...

07 July 2014 8,060 13 View

Can you help with my problems during ChIP sonication procedure?

I and my colleagues are trying to master the methodology of chromatin immunoprecipitation (ChIP) of mouse skeletal muscle. We performed ChIP protocol and used sonication to shear the chromatin to...

07 July 2014 2,774 3 View

Problem with agarose gel electrophoresis voltage and time. Can anyone help?

I want to run DNA fragments around 200-300 bp. In your opinion, is it ok to use 1.5% agarose, and what do you suggest about voltage-intensity and time? The gel dimension is 8 x 10 cms. I have...

06 June 2014 3,113 14 View

Does anyone have experience with cooled gel-electrophoresis?

Usually, voltage at electrophoresis is pushed as far as possible without producing smiling gels due to the gel heating up. I am currently playing with the thought of building an electrophoresis...

06 June 2014 5,721 10 View

How can I synthesize a DNA ladder using lambda DNA? Would like opinions on what's worked for you?

I want to create my own DNA ladder for gel electrophoresis in order to save money. Has anyone any recommendations on particular enzymes to use or the most validated methodology? We have run a few...

06 June 2014 9,734 5 View

What is the value of the dielectric constant/relative permittivity of electrophoresis gel?

I am trying to simulate (in Comsol) electric field through an electrophoresis system and need an approx. value for the dielectric constant/relative permittivity of any kind of gel (like PAGE or...

06 June 2014 2,884 1 View

Does anyone have a good protocol for running Native PAGE?

My protein has a PI in the physiological pH and is in the phosphate buffer, pH 6.0. I would like to check if the protein is in the monomeric state.

06 June 2014 4,117 4 View

What is wrong with my gel electrophoresis?

I am having trouble running a gel electrophoresis on plasmid DNA. I was having acceptable results until a couple weeks ago when my bands started smearing together. I have tried new running buffer...

06 June 2014 7,141 7 View

Should we use stacking and resolving gel in PAGE for RNA?

I want to separate RNA on PAGE. Should I use stacking or resolving gel?

05 May 2014 4,209 1 View

Can anyone help with a problem regarding agarose gel electrophoresis?

While doing agarose gel electrophoresis, the bands are either very light or not visible despite the amount of good DNA quality and quantity. I am using 1.5ul EtBr for 30ml of gel. I have already...

05 May 2014 6,286 11 View

Can anyone help with sample loading for quantitative proteomics?

In my proteomic analysis, I have tried to a relative quantitation of proteome using 1D Gel LC/MS following by a widely know spectral counting method called as Normalized spectral abundance factor...

04 April 2014 962 3 View

Can anyone suggest a gel system which will reliably resolve >700 kDa bands?

I'm studying the dimerization of a high-weight protein (~350 kDa), which means that I need to be able to resolve both a 350 kDa band and a 700 kDa on a gel. I've gone through several different...

04 April 2014 6,038 4 View

Is the "HMW Native Marker Kit" from GE Healthcare suitable for BN PAGE?

I am looking for a molecular weight marker that can be used for BN PAGE, and I am wondering if the product metioned above gives nice sharp bands. I am also open for any other suggestions you might...

04 April 2014 6,185 3 View

Can we precipitate protein from 7M urea and 2M thiourea containing buffer using acetone/TCA?

For 2D

04 April 2014 4,156 5 View

Has anyone tried to add TMAO (or PEG20K) in gels for electrophoretic separation of proteins (non-denaturing conditions)?

The idea is to visualize high-order oligomers of a protein which can self-associate but normally exists as small oligomers. Is it OK to use native-PAGE and gels containing TMAO? Which TMAO...

04 April 2014 1,152 0 View

How do proteins get their charge in native gel electrophoresis? Can any one explain in detail?

I'm going to do native page and i just want to know its principle completely.

04 April 2014 9,763 7 View

Does anyone have experience with Bolt Bis-Tris Plus gels from Life Technologies?

Can the MOPS running and transfer buffers that go with the older Nupage Bis-Tris gels be used with the Bolt gels? If not, does anyone have the recipe for the Bolt running and transfer buffers?

04 April 2014 475 1 View

Where can I sequence my protein spots?

Can somebody help me to inform any company providing for protein spots sequencing service (MALDI-TOFF or LC MS/MSS Platform)? We have some interesting protein spots that need to be further...

04 April 2014 4,320 2 View

What is the probable cause of particular failure at the Pulsed field gel electrophoresis?

Although we can complete our protocol with other strains, we confront the same problem over over again with same patient. Picture of PFGE is attached. The left one belongs to another patient and...

04 April 2014 1,886 2 View

Can a normal room temperature camera in gel doc produce quantitative images?

The BioRad geldoc system comes with two types of cameras, one CCD camera which operates at room temperature and a super cooled CCD camera (-30C). Can we use images taken from the room temp camera...

04 April 2014 2,116 4 View

Is there anyone that has used PDQuest software?

I am trying to upload my 2-D gel images to PDQuest software virgin 8.0.1, but the software refuses them. Despite that at first it asked me to upload them in Grey scale, which I have already done,...

04 April 2014 5,849 3 View

2D gel Oriole Staining Background Issue, can anyone help?

I recently ran a 2D gel electrophoresis, however my spots were a wee bit faint, so i increased the exposure of the image. This resulted in the appearance of small pixels (dirty spots?) across my...

04 April 2014 4,890 2 View

Is there anyone that has used PDQuest software?

I am trying to upload my 2-D gel images to PDQuest software virgin 8.0.1, but the software refuses them. Despite that at first it asked me to upload them in Grey scale, which I have already done,...

01 April 2014 6,228 3 View

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