TAE Buffer and pH

[jrd210]

I am somewhat confused. Being new to PCR and electrophoresis I am mixing up TAE in 50-1 strength with Glacial Acetic acid and EDTA  added in correct proportions and finding the end pH to be far more acidic than it is documented on most sites. Some sites say do not test your mix after making up TAE and just make sure the mix is correct.

Others say it must get to 8.3, well I tried adding NaOh 1M and found that I seemed to need a great deal to bring it close to 8.3 and have actually run through my current supply of NaOh.

How much of an issue is getting this pH to around 8.0???  I would welcome some suggestions.

[Dakota Hamill]

How did you make it?  What and how much did you mix together?

I've never pH d my tae and its always worked. 

--
-- You received this message because you are subscribed to the Google Groups DIYbio group. To post to this group, send email to diy...@googlegroups.com. To unsubscribe from this group, send email to diybio+un...@googlegroups.com. For more options, visit this group at https://groups.google.com/d/forum/diybio?hl=en
Learn more at www.diybio.org
---
You received this message because you are subscribed to the Google Groups "DIYbio" group.
To unsubscribe from this group and stop receiving emails from it, send an email to diybio+un...@googlegroups.com.
To post to this group, send email to diy...@googlegroups.com.
Visit this group at https://groups.google.com/group/diybio.
To view this discussion on the web visit https://groups.google.com/d/msgid/diybio/1203613e-94f5-4010-8af6-489c3608cc8a%40googlegroups.com.
For more options, visit https://groups.google.com/d/optout.

[Cory Tobin]

> I am mixing up TAE in 50-1 strength with Glacial
> Acetic acid and EDTA added in correct proportions

I hope you're adding tris-base too. Without the tris-base the pH
would be very low due to the acetic acid and would require a lot of
NaOH to bring the pH above 8. If you've added the correct components
in the correct proportions you shouldn't have to adjust the pH, even
if you start with very acidic water.

-cory

[Dakota Hamill]

Yeah forgetting your base would do it!

--
-- You received this message because you are subscribed to the Google Groups DIYbio group. To post to this group, send email to diy...@googlegroups.com. To unsubscribe from this group, send email to diybio+un...@googlegroups.com. For more options, visit this group at https://groups.google.com/d/forum/diybio?hl=en
Learn more at www.diybio.org
---
You received this message because you are subscribed to the Google Groups "DIYbio" group.
To unsubscribe from this group and stop receiving emails from it, send an email to diybio+un...@googlegroups.com.
To post to this group, send email to diy...@googlegroups.com.
Visit this group at https://groups.google.com/group/diybio.

To view this discussion on the web visit https://groups.google.com/d/msgid/diybio/CAC46JssP5DFfUQY-C4kKFbn4GtmY2ccazcLr7VJ%3DOAa57TnE9g%40mail.gmail.com.

[Tom Randall]

Look at the Trizma buffer table in this link, very useful if making from scratch:

http://www.sigmaaldrich.com/life-science/core-bioreagents/biological-buffers/learning-center/buffer-reference-center.html