Simple and efficient subtractive hybridization screening. |
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Authors: | M R Herfort A T Garber |
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Institution: | Department of Medical Biochemistry, University of Calgary, Alberta, Canada. |
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Abstract: | A method of subtractive hybridization screening is presented that does not rely on the availability of large amounts of poly(A)+ RNA. The usefulness of this method is demonstrated by the isolation of cDNA clones that are unique or present at enhanced levels in cDNA libraries derived from a psoriatic skin cell line. A very high level of enrichment for rare cDNA sequences is possible as shown by the isolation of cDNA clones present at a frequency of less than one in one hundred thousand (0.001%). This method is particularly useful in experimental systems where the amount of RNA available is limited. |
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