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. 1994;1(5):245-50.
doi: 10.1093/dnares/1.5.245.

A PCR-mediated method for cloning spot DNA on restriction landmark genomic scanning (RLGS) gel

H Suzuki  1 , J Kawai, C Taga, N Ozawa, S Watanabe
Affiliations
Free article

A PCR-mediated method for cloning spot DNA on restriction landmark genomic scanning (RLGS) gel

H Suzuki et al. DNA Res. 1994.
Free article

Abstract

A PCR-mediated direct cloning for target spot DNA from RLGS gel has been established. The method consists of PCR amplification of adaptor-ligated spot DNA fragments without excluding similar-sized DNA fragments co-localized on RLGS gel, and following selective ligation with the NotI-dT vector. Applying this method, we have successfully cloned several DNA fragments derived from target spots whose intensities change developmentally due to DNA methylation in the telencephalon of C3H/HeN mice. Since only a few micrograms of total DNA is sufficient for our spot cloning, our method may be highly useful when the total DNA sample prepared for cloning is limited.

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