Please help me ...
which is the best solvent for dissolve and prepare DNA standard solution for estimation .
You can dissolve your DNA in sterile water. Its pH should be above 8.0, If you use Tris-HCl including EDTA, that may cause problem in further experiments, e.g. PCR reaction, protein binding assays. EDTA is a strong chelator, it will chelate all Mg++ that binds to DNA bases and are necessary for reactions during PCR or any other. On the other hand you can use Tris-HCl EDTA (TE) buffer, but then it will need more Mg++. Usually commercial kits suggest to store DNA in water, or Tris-HCl 10 mM, pH 8. I dont know what kind of DNA you are using? and what kind of analysis you do?, but the poor results might be also due to improper dissolving DNA in buffer. Hope it help you!
TE with PH 7.8- 8 is considered best and it increases the storage life of DNA too. You can also use sterile PCR grade water to dissolve and estimate your DNA.
thank you sir.
but ,
we used TE buffer with 7.6 ph. but shows poor results.
can you please give the exact composition of TE buffer.
You can dissolve your DNA in sterile water. Its pH should be above 8.0, If you use Tris-HCl including EDTA, that may cause problem in further experiments, e.g. PCR reaction, protein binding assays. EDTA is a strong chelator, it will chelate all Mg++ that binds to DNA bases and are necessary for reactions during PCR or any other. On the other hand you can use Tris-HCl EDTA (TE) buffer, but then it will need more Mg++. Usually commercial kits suggest to store DNA in water, or Tris-HCl 10 mM, pH 8. I dont know what kind of DNA you are using? and what kind of analysis you do?, but the poor results might be also due to improper dissolving DNA in buffer. Hope it help you!
I am sorry i made mistake to write the composition of TE buffer its 10mM Tris-HCL and 0.1 mM EDTA. Birendra Singh is right.
@Nandheesh...
1. Dissolve your main stock in TE buffer as mentioned above for longer usage.
2. For working stock go for sterile/nano pure water from main stock because this will eliminate your next problems like PCR etc......
Mean while let me know how are you going to tell that your DNA is poor quality?????
Based on gel image ???? or quantifying it on spectrophotometer/nanodrop instrument??? let me know if you have results.....
good luck...
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