9 replies to this topic

[noyara]

Hi Friends, How do you do?

I wander which buffer is more popular in gel elctrophorsesis? for me , i used both to compare between them, i found TBE produce more clear and nice bands than TAE, but other labs used TAE much more than TBE,
So, which one do you use for your gel electrophoresis?

Please vote..     

Thank you in advance
Noor

[Ameya P]

Your First voter Noyara  

Technically, I use 0.5X TBE buffer. All my PCR products are under 1000bp and TBE is good when it comes to resolving smaller fragments.

[noyara]

gt_ameya, on 16 April 2011 - 02:42 AM, said:

Your First voter Noyara  

Technically, I use 0.5X TBE buffer. All my PCR products are under 1000bp and TBE is good when it comes to resolving smaller fragments.

Welcome ameya,
Why used 0.5 X TBE instead of 1X TBE?

[Ameya P]

I dont quite know, but it works.... on the long run, saves you money  

[hobglobin]

Lithium Borate is the best...fast, high resolution and cheap...

[grvsomani]

i use 1X TAE. and use to amplify 1 kb to 3 kb product with no problem. my ladder also resolve nicely. i use invitrogen agarose.

[noyara]

hobglobin, on 16 April 2011 - 05:03 AM, said:

Lithium Borate is the best...fast, high resolution and cheap...

Hi,
I think LB used for small fragment less than <3Kb , is it work for large fragments as well?
and i wander at which Voltage you run the gel, is it(5-50 V/cm)? so for 8 cm set the V at 400 V ?!! and the time is it finished within 10-15 min?!!

normally for 1X TBE i used 80 V,for 45 -60 min.

It will be really amazing if can finish within 10 min !!

[lab rat]

gt_ameya, on 16 April 2011 - 02:42 AM, said:

Your First voter Noyara  

Technically, I use 0.5X TBE buffer. All my PCR products are under 1000bp and TBE is good when it comes to resolving smaller fragments.

us too. we reuse gel and buffer several times, which also saves money.

[hobglobin]

noyara, on 16 April 2011 - 08:09 AM, said:

hobglobin, on 16 April 2011 - 05:03 AM, said:

Lithium Borate is the best...fast, high resolution and cheap...

Hi,
I think LB used for small fragment less than <3Kb , is it work for large fragments as well?
and i wander at which Voltage you run the gel, is it(5-50 V/cm)? so for 8 cm set the V at 400 V ?!! and the time is it finished within 10-15 min?!!

normally for 1X TBE i used 80 V,for 45 -60 min.

It will be really amazing if can finish within 10 min !!

with larger fragments there is no real difference, but it works, only quite faster, i.e. a gel is finished in 10-20 minutes. But our power supplies won't make 400V, so 200V is maximum for smaller DNA (we work mostly with sizes between 100-500 bp).

[Adrian K]

lab rat, on 16 April 2011 - 08:22 AM, said:

gt_ameya, on 16 April 2011 - 02:42 AM, said:

Your First voter Noyara  

Technically, I use 0.5X TBE buffer. All my PCR products are under 1000bp and TBE is good when it comes to resolving smaller fragments.

us too. we reuse gel and buffer several times, which also saves money.

Hi lab rat and Ameya,
I too use 0.5X TBE as well... the gel buffer was reuse till it looks very dirty before we discard it... lolx....