TBE vs TAE for agarose gel electrophoresis – (Aug/06/2006 )


TBE vs TAE for agarose gel electrophoresis – (Aug/06/2006 )

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Hi,

I would just like to know, is there any major difference between TBE buffer and TAE buffer for agarose gel electrophoresis. Plus, is there any different between 0.5x TBE and 1.0x TBE in the results?

Thank you.

-Aquilon-

QUOTE (Aquilon @ Aug 7 2006, 04:39 PM)
Hi,

I would just like to know, is there any major difference between TBE buffer and TAE buffer for agarose gel electrophoresis. Plus, is there any different between 0.5x TBE and 1.0x TBE in the results?

Thank you.

Essentially, no to both of your questions (I think that Maniatis, or Sambrook (=Maniatis edition 2) go into the difference, but it’s not significant enough to lose sleep over it). 0.5x TBE is just less buffered. As long as your gel and the buffer are the same strength.

-swanny-

Ok, thanks.. Just wondering..

-Aquilon-

TAE is somewhat less fussy about extraction of the DNA from bands cut from the gel. TBE requires some additional chemistry during the extraction process. Any of the gel extraction kits will describe this in detail.

-phage434-

TBE is supposed to improve the ‘sharpness’ or consistency of the migrating bands, but I never really saw a difference.

As phage434 mentioned, its a little more complicated to extract from – I’d stick with TAE if there is a choice.

-vasussci-

I believe TAE has less ‘buffering capacity’ than TBE. That means you can reuse TBE atleast 3 times while TAE can’t be used more than once. The bands start getting smeary.

-lotusgirl-

I would say that is untrue, based on my experience. I have not used TBE in many years, but I use TAE all the time and re-use it, usually 3-4 times

-aimikins-

i havent had problem extracting DNA from the gel made using TBE. My lab uses it and our extraction procedue is not different to what others use for TAE.

I have reused TAE and TBE both for many times without any problem.

-scolix-

If you make the agarose gel in TAE buffer and run the gel submerge in TBE buffer, the gel will not run.
I had try it and got into trouble ph34r.gif .

-Minnie Mouse-

0.5X TBE is having less buffering capacity than1 X so 1X is minimum for running a gel and diff b/n TBE TAE is what i read is that when u r running a gel it generates huge amount of heat so buffer will exhaust some thing after some time at high volts compare to TBE TAE doesnt do this job so it is preferable, and
if u observe onething after some time u can see the evaporation on the top side of the tank ok this indicates evaporation takes place due to heat generation

-Add colour 2 ur life-

in our lab we use 0.5x … no problems there, but you can run agarose gels quite fast, as the ionic strength and hence the resistance is lower. Means less heat, no melting of gel at higher power

-Kersten-

I have to agree with scolix, I have never altered my gel extraction protocol for TBE gels.

The only thing I ever came across was when gel extracting a transgenic cassette for subsequent microinjections the protocol insisted on using TAE gels and to not use TBE but the protocol never went into the theory behind why.

-JPStewart-

Use SB, no tris, just sodium hydroxide and boric acid, so cheap and easy to make as well as good resolution and can run gels fast.

-bob1-

One of my lab mates is running alot of electrophoresis, in a few days, we notice that the tank is very hot, but the gel is still fine then one day smoke came out from the electrophoresis machine (not the tank) and it cannot be use anymore. We suspected it is the buffer that he made was inaccurate. Is there anyway to check it?

-Aquilon-

We use 0.5x TBE regulary and 1x TBE for long (day/night) electrophoresis, because of the believed better buffering capacity of TBE. 1x TAE is used in our lab for gel extractions because we are used to and some people claims it gives higher yield, but I never seen any proof to that.
I would personally say, that bands look a bit sharper in TBE (0.5) than in TAE.

-Trof-

Cambrex company has”A handbook for gel electrophoresis” in their website. According to this handbook,

TAE is used for DNA recovery and electrophoresis of large (>12 kb) DNA. TAE has low buffering capacity and recirculation may be necessary for >6 h of electrophoresis.

TBE is used for small (<1 kb) DNA and shows increased resolution of small DNA. TBE shows decreased DNA motility and has high buffering capacity, therefore recirculation is not needed for extended run times.

For electrophoresis, both buffers work for me. For DNA extraction I am using gel extraction kits and they are designed to extract and purify DNA from agarose gels in TAE and TBE buffer. I could not see a difference in the purity or amount of extracted DNA when I used any of these two buffers. Probably, for long running times or repeated use, TBE is better. Repeated use of TAE buffer results in excessive heat and failure of electrophoretic equipment.

-branmorn-

 

QUOTE (branmorn @ Aug 18 2006, 02:48 PM)
Cambrex company has”A handbook for gel electrophoresis” in their website. According to this handbook,

TAE is used for DNA recovery and electrophoresis of large (>12 kb) DNA. TAE has low buffering capacity and recirculation may be necessary for >6 h of electrophoresis.

TBE is used for small (<1 kb) DNA and shows increased resolution of small DNA. TBE shows decreased DNA motility and has high buffering capacity, therefore recirculation is not needed for extended run times.

For electrophoresis, both buffers work for me. For DNA extraction I am using gel extraction kits and they are designed to extract and purify DNA from agarose gels in TAE and TBE buffer. I could not see a difference in the purity or amount of extracted DNA when I used any of these two buffers. Probably, for long running times or repeated use, TBE is better. Repeated use of TAE buffer results in excessive heat and failure of electrophoretic equipment.

Would you please provide us with the website link? 

-Minnie Mouse-

 

QUOTE (Minnie Mouse @ Aug 21 2006, 08:00 AM)
QUOTE (branmorn @ Aug 18 2006, 02:48 PM)
Cambrex company has”A handbook for gel electrophoresis” in their website. According to this handbook,

TAE is used for DNA recovery and electrophoresis of large (>12 kb) DNA. TAE has low buffering capacity and recirculation may be necessary for >6 h of electrophoresis.

TBE is used for small (<1 kb) DNA and shows increased resolution of small DNA. TBE shows decreased DNA motility and has high buffering capacity, therefore recirculation is not needed for extended run times.

For electrophoresis, both buffers work for me. For DNA extraction I am using gel extraction kits and they are designed to extract and purify DNA from agarose gels in TAE and TBE buffer. I could not see a difference in the purity or amount of extracted DNA when I used any of these two buffers. Probably, for long running times or repeated use, TBE is better. Repeated use of TAE buffer results in excessive heat and failure of electrophoretic equipment.

Would you please provide us with the website link?

Hi Minnie
here is the book
http://www.cambrex.com/Content/Documents/T…eBook_FINAL.pdf 

-spanishflower-

 

QUOTE (spanishflower @ Aug 20 2006, 05:59 PM)

Thank you Spanishflower. smile.gif 

-Minnie Mouse-

 

QUOTE (Aquilon @ Aug 6 2006, 10:39 PM)
Hi,

I would just like to know, is there any major difference between TBE buffer and TAE buffer for agarose gel electrophoresis. Plus, is there any different between 0.5x TBE and 1.0x TBE in the results?

Thank you.

TBE is supposed to produce a sharper band, but i agree with the person who said the difference in visual quality isn’t great.

we use SYBR green and reuse 1x TAE buffer and the gel itself many, many times (at least 10), just topping it up to account for evaporation. 

-MingTea-

TBE vs TAE for agarose gel electrophoresis – (Aug/06/2006 )

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hi!!

i just want to noe if anyone can share with me the recipe for 40XTAE buffer with me. i juz want to compare with mine coz after gel electhrophoresis, my gel n the tank became hotter than when i use TBE buffer before. juz wanna check if i did messed up in the recipe. thanks!!

-icemizu-

 

QUOTE (lotusgirl @ Aug 7 2006, 03:23 PM)
I believe TAE has less ‘buffering capacity’ than TBE. That means you can reuse TBE atleast 3 times while TAE can’t be used more than once. The bands start getting smeary.

I have used TAE atleast three times and i had no probs.. TBE has better resolution on DNA gels, and for DNA it is better to use 1X conc. for protein 0.5 X is okay
good luck
she 

-teshee-

 

QUOTE (Aquilon @ Aug 6 2006, 11:39 PM)
Hi,

I would just like to know, is there any major difference between TBE buffer and TAE buffer for agarose gel electrophoresis. Plus, is there any different between 0.5x TBE and 1.0x TBE in the results?

Thank you.

Hi,

Yes there is a difference between TBE and TAE that is their electroconductivity

In case of agarose gel electrophoresis it is better to use TBE

0.5X TBE has half the empirical amount of Tris, boric acid and EDTA ahere as in case of the 1.0X TBE it contains the empirical amount of the buffer constituents. 

-abhishek.harichandan-

I have seen from the net somewhere that its better to avoid TBE in extraction of band if you are to use in cloning or synthesis due to the borate salt and use TAE instead. I notice this when I use platinum taq for amplification and the gel I run tapered at the end but not present when I rerun using TAE gel and buffer.

but we use most of the time TBE….

-friley-

in our lab, we use 0.25X TBE in our tanks and gels (2-2.5% agarose).
our products are around 200 kb (+/- 100)
we change (refresh) these every monday.
we run approx 25 gels 1hr each at 200V (constant voltage) per week in each of our tanks.
we have no problems, though we do at times need to topup the buffer due to eveaporation.

hope that helps.

-wiklendt-

 

QUOTE (lotusgirl @ Aug 7 2006, 02:23 PM)
I believe TAE has less ‘buffering capacity’ than TBE. That means you can reuse TBE atleast 3 times while TAE can’t be used more than once. The bands start getting smeary.

I use TAE all this while. everyone in my lab use TAE… we reuse many many many times, for whole week… still OK. 

-sanjiun81-

I think I agree with friley, in my lab also we use TAE for cloning whereas in other neighbouring labs I’ve seen that people use TBE instead of TAE, I dint know why but now I guess bcoz they do it for polymorphism. [smile.gif]

-mysterious-

I partialy solved my problem with increased heating of the TAE buffer, because I run it in the fridge smile.gif
I can use a slight higher voltage and the bands look better, IMO.

-Trof-

I’ve noticed that if I allow my gels to set in the fridge (and get really cold) then the bands run sharper. It’s particularly good for running restriction digests because they have a higher salt content so they run hotter and produce blurry bands (especially at 250v+)

-Zouden-

I use .5X TAE all the time and have gotten excellent results from PCR/RT-PCR products. For better signal I sometimes add 2~4uL EtBr to the + end buffer solution in the apparatus.

-Rixin-
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