I need to dilute GAPDH primer for PCR. In the brochure they mentioned TE buffer pH 7.5 as the diluent. But, I have TE buffer 8.00 at my hand now. Is it OK or may cause any deviation with TE 8.00?
Hi there,
pH8 is definitely better for DNA as its depurination is an acid catalysis whereas pH7.5 is better for RNA because RNA degradation is base catalyzed.
You can dilute your lyophilized primers in autoclaved MilliQ (double distilled) water. I always dilute it like this only and it works well for any reactions.
Regards
Hi Monirul, I use GAPDH and many other primers for PCR and always dilute them in nuclease free water.
If you resuspend/dilute them in water, they can be used for a variety of reactions, so that's what we do, and we keep them frozen in well-annotated aliquots so that they can still be found and used years afterwards (if necessary).
I do not preferto add buffers. DNAse and RNAse water is most prefereable in our lab.
TE buffer at pH 8 is completely okay to dilute primers. You can also use DNAse water for dilution.
We routinely make primer stock (100uM) in TE pH 8.0, but dilute it to 10uM for for use, with water. The presence of too much EDTA may chelate MgCl2, affecting the PCR reaction, therefore minimizing the EDTA is advisable.
- Badges
- Science topic
- Similar topics
- Biological Science
- Biochemistry
- Biochemical Techniques
- Analytical Chemistry
- pH