I have seen in lab that my colleagues used different concentration of NaCl for CTAB preparation as per their tissue requirement. Is there any body who can update me on the required concentration of ingredients for CTAB preparation for my blood samples? Beside this also tell me why we use different salt concentrations for different tissues. What will be the effect of salt?
but what is the established protocols? kindly provide me and tell me most important question of salt concentration effect.
I assume you use the CTAB extraction where you use chloroform for separation of proteins. As CTAB is a detergent, it will emulsify your aqueous phase in the organic phase for some part. If you raise the ionic strength in the aqueous phase with the addition of salts, you will break the emulsion and the sample is more likely to separate, increasing the yield of aqueous phase with DNA.
I think you may want to increase the salt concentration in your aqueous phase if you are handling with very small sample sizes, but honestly... Extraction protocols are not perfect, some numbers in there are just that way for arbitrary reasons and can be varied without much consequences.
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