For my experiment I have to extract DNA from frogs. I wanted to use a CTAB method but I have a RNA contamination. How can I remove this whitout using RNase or other enzymes? It has to be as cheap as possible!
Phenol:chloroform is cheap and efficient:
H.E. McKiernan, P.B. Danielson, in Molecular Diagnostics (Third Edition), 2017
21.3.1 Organic (Phenol–Chloroform) Extraction
Organic (phenol–chloroform) extraction uses sodium dodecylsulfate (SDS) and proteinase K for the enzymatic digestion of proteins and nonnucleic acid cellular components (Fig. 21.4). A mixture of phenol:chloroform:isoamyl alcohol (25:24:1) is then added to promote the partitioning of lipids and cellular debris into the organic phase, leaving isolated DNA in the aqueous phase. Following centrifugation, the aqueous phase containing the purified DNA can be transferred to a clean tube for analysis. DNA can also be recovered and concentrated from the aqueous phase by ethanol precipitationor through the use of a centrifugal filter unit (i.e., Vivacon® or Amicon® devices), which allows for additional purification and concentration of the DNA in the samples (Koons et al., 1994). Organic extraction recovers double-stranded DNA and was required for early RFLP methods. While this method remains one of the most reliable and efficient, it is also very time-consuming, uses hazardous chemicals, and, because of the greater hands-on effort and multiple tube transfers involved, introduces increased opportunities for contamination and sample mishandling (Köchl et al., 2005).
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