Currently we use novex 4-12% bis-tris gels from life technologies. We can run them at 200V in MES buffer, which takes 35 minutes to run.
I was wondering if there was a protocol to hand make gels that would run faster than conventional hand made gels (i.e. less than an hour).
Does anyone know what is in the composition of these gels that makes them run fast and give good resolution?
The gel run time depends on the used buffer system (gel and running buffer), the geometry of the gel system and the applied power. While using smaller gels (mini gels) reduces the duration of the gel run, increasing the power often heats up the gels that can decreased quality of the separation.In my opinion, 35 min for SDS-PAFE is already very fast.
Best regards
I agree with the above person. 35 minutes is already really fast. You can lower the overall percent of the gel, aka hand pour a 6% gel that will run pretty fast. But really there is not a huge benefit to running a gel faster. Sometimes you sacrifice quality for speed.
Applying higher voltage will make it run faster but will also produce a lot of heat (gels have high resistance). You can try running in cold running buffer inside an ice-water bucket (don't wet your connections) and increase voltage. Try with 'cheap' samples first.
Yoram
I agree with the people above. 35 minutes is really fast. With handmade minigels (8x10 cm) the runs usualy take around 90 minutes So I don´t think you could make it faster without compromising resolution or overheating the gel.
Patrick
35 min is already faster. So if you go further increase you may not get the resolution of bands that you really require.
I agree with the above opinions of the colleagues. Too fast for a quality resolution. Try some points: Optimize the gel PA % for your band size resolution; Apply a lower voltage and Wait a little more... I hope it works!
Why do you want to run the sample faster? I think you're about at the limit with what you're doing - you might be able to run the gel faster in the cold room with a recirculating cooling system of some sort, but you will probably lose resolution.
If you want faster results, and if your lab has money, something like the Agilent Bioanalyzer (lab-on-a-chip, microfluidic analysis) will give you fast results, but it's much more expensive than a PAGE setup.
Use Creiterion Gels from Bio-Rad - running time is 20 min in their buffer, with brilliant resolution.
http://www.bio-rad.com/en-us/product/criterion-precast-gels/criterion-tgx-stain-free-precast-gels
If you use the stain free gels, you don't even have to do comassie - you will need to buy an imager that uses A280 absorbance to scan your gels, but the scanning is done in 2.5 min. If your protein does not have tryptophans, you can stain these gels in Comassie or any other stain that you use. Stain free gels (20 min running plus 2-3 min imaging) has changed my life - saves a ton of time.
for DNA in PAGE GEL , what is best ratio for electrophoresis with PCR product in gel??
U= ?
I=?
P=?
Time??
can anyone tell me please.
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