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Hi,
I am trying to prepare the Turbo DNA-free second digest 10X buffer and was hoping someone might be able to help me with some questions. The protocol (from life technologies website) is:
"200 mM Tris-HCl pH 7.5, 100 mM MgCl2, and 5 mM CaCl2, prepared with nuclease-free H2O. The final pH of the solution should be 7.5. If the pH is not 7.5, add NaOH or HCl as necessary, but do not introduce more than 50 mM of total monovalent salt to the 10X buffer when adjusting the pH in this way."
I was wondering how I can keep the final buffer RNase free as adding each component to nuclease free water will probably introduce small amounts of RNase because each media component would not be 100 % pure. As well, I'm wondering how to measure the pH as the pH probe will also be contaminated. Can I maybe wipe it down with 0.1 M NaOH and let it sit for a couple minutes before use?
Thanks
I am trying to prepare the Turbo DNA-free second digest 10X buffer and was hoping someone might be able to help me with some questions. The protocol (from life technologies website) is:
"200 mM Tris-HCl pH 7.5, 100 mM MgCl2, and 5 mM CaCl2, prepared with nuclease-free H2O. The final pH of the solution should be 7.5. If the pH is not 7.5, add NaOH or HCl as necessary, but do not introduce more than 50 mM of total monovalent salt to the 10X buffer when adjusting the pH in this way."
I was wondering how I can keep the final buffer RNase free as adding each component to nuclease free water will probably introduce small amounts of RNase because each media component would not be 100 % pure. As well, I'm wondering how to measure the pH as the pH probe will also be contaminated. Can I maybe wipe it down with 0.1 M NaOH and let it sit for a couple minutes before use?
Thanks