How to make 400 mM Tris-HCl pH 9.0 + 10 mM MgCl2

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CSUBARCODER
CSUBARCODER's picture
How to make 400 mM Tris-HCl pH 9.0 + 10 mM MgCl2

 Hi all.

I am trying to make extra sequencing buffer which is typically used with the BDRR V3.1 Kit for cycle sequencing.

I am told this is what it is:

400 mM Tris-HCl pH 9.0 + 10 mM MgCl2
 
Can someone give me step by step instructions on the best way to make this solution.

Thanks.

Brian

Ivan Delgado
Ivan Delgado's picture
Hi Brian,

Hi Brian,

You should be able to get the information you need here. In that page they explain how to make 5x replacement buffer, what you are looking to make, plus gives multiple links to other websites that talk about this buffer. 

Good luck!

CSUBARCODER
CSUBARCODER's picture
You Rock Ivan.  Thanks!

You Rock Ivan.  Thanks!

cibertech
cibertech's picture
Hello CSUBARCODER/ Ivan

Hello CSUBARCODER/ Ivan

I want to reply to this post, altough I run the risk of doing some publicity here, as I am not impartial.
There are vendors who have alternative reagents for these machines. One of them is geographicaly very close to Ivan, actually, and probably he already knows it. 
With these reagents I'm refering not only to the Buffer CSUBARCODER mentions above, but also the polimers for the machines. And even polymers which have the exact same formulation as the manufacturer, in the case of the 4 & 6 versions. 
This is especially important now, as the manufacturer will phase out these polymers, so it might be interesting for you to chek out.

As I cannot post any publicity here, you can send me a direct message if you need more information.

I hope this was usefull.

Best regards

Ezequiel Coelho

Ivan Delgado
Ivan Delgado's picture
Hi Ezequiel,

Hi Ezequiel,

Thank you for your message. It is a valid point. 

Brian: I would contact Ezequiel by private message (PM) and ask about the vendors he is referring to.

Cheers

cibertech
cibertech's picture
 Hello Ivan

 Hello Ivan

Thanks for the reference.
By the way, may I ask you what kind of machines do you use in your lab? 3730's?

Best regards

Ezequiel

Ivan Delgado
Ivan Delgado's picture
Hi Ezequiel,

Hi Ezequiel,

Anytime. 

Actually we do not have capillary gel electrophoresis instruments in my lab. I used to work at companies that did that kind of work (that is where my experience comes from). At those companies we used everything from ABI 310s and 377s to ABI 3730xls.

Best