Plz refer the following links. Hope this should help
are tris Cl and tris HCl are same?
i think it is just the usage of PH dependent buffer solution. HCl makes the Tris PH value under 7. I hope this could help....
For Tris-base you add HCl for adjusting pH
most of the times I prepare tris buffer and adjust to ph 7.2.. after filtration I have observed increase in ph of about 0.2 to 0.3.. how can I come over with this problem?
You can calculte how to prepare tris hcl from tris base here:
Dear experts , what i observed that when i used Tris-base to prepare TAE/TBE buffer, the resulting pH was around 8.5, however when i used Tris-HCL the resulting pH was below 4.0.
In order to decrease the pH i used to add HCL and the TAE/TBE buffer works well.
However using Tris-HCl (pH 4.0) i need to add NaOH . when i check this buffer for electrophoresis , it gives very low voltage ( even not able to adjust by adjustable power pack) .
Can anyone suggest me some solution for the above problem
"it gives very low voltage" This may be because your power supply has an "amperage" limiting feature. When you adjusted with NaOH, you added much more sodium ion to the solution, so at the same voltage, you are getting much more current. Did you monitor the current (amps)? For a fixed voltage, the current is a function of the ion concentration, so if there are more ions present, the current (amps) will be higher. If your power supply is "amp-limited", you can only adjust the voltage up to the point where the amperage is maximal. After that, no adjustment will increase the voltage and the current will remain at the maximum value. And since higher current also means higher heat and higher heat will melt your gel, you don't want to run at higher amps anyway.
Actually, the best solution for agarose gel electropheresis is not to use Tris "buffer" at all, but rather a simple "electroconductive medium" described in the attached papers.
A simple recipe for a 20XSB stock useful for most RNA and DNA applications, and requiring no further pH adjustment is:
For 4 liters 20X SB stock
NaOH 32 gram
Boric Acid 200 gram
Tap distilled/deionized water to 4 liter
Thank you David.
How do I adjust the pH of Tris-HCl to 8.8, if I mix Tris-HCl directly with DI water, instead of Tris base and HCl ?
SB Buffer is easy to prepare and works better than TAE and TBE.
SB buffer Composition-20X (1L)
Boric acid - 50 grams
Mix properly and use 1X as working Buffer.
Thank you, Dr. David for your valuable suggestion.
Lalan Kumar Arya
Madurai Kamaraj University
Florida International University
University of the Punjab
Central Manchester University Hospitals NHS Foundation Trust
Universiti Sains Malaysia
Umm Al-Qura University
Abdulkareem Ali Alanezi
University of the Sciences in Philadelphia
Federal University Wukari, Nigeria
Research Genetic Cancer Centre Ltd