Cloning genes from a library using a clustering strategy and PCR.
A new polymerase chain reaction (PCR)-based method is described for the isolation of clones of interest from a library when only part of a sequence is available. In actuality, this occurs with many genomes that have been partially sequenced using a random strategy. The method presented here, discriminating clusters by PCR (DCbyPCR), is a nonradioactive and improved alternative to colony hybridization.
Díaz Prado
S
S
Department of Molecular and Cell Biology, University of A Coruña, Campus da Zapateira s/n, 15071-A Coruña, Spain.
Tarrío
N
N
Cerdán
M E
ME
González Siso
M I
MI
D003001:Cloning, Molecular; D004926:Escherichia coli / Q000378:metabolism; D004996:Ethidium / Q000494:pharmacology; D015723:Gene Library; D005821:Genetic Techniques; D007364:Intercalating Agents / Q000494:pharmacology; D007716:Kluyveromyces / Q000378:metabolism; D009693:Nucleic Acid Hybridization; D010957:Plasmids / Q000378:metabolism; D016133:Polymerase Chain Reaction; D012685:Sepharose / Q000737:chemistry
D007364:Intercalating Agents; D012685:Sepharose; D004996:Ethidium