A rapid reliable method to use specific probes labelling polymerase chain reaction (PCR) products.
Polymerase chain reaction (PCR) has allowed highly sensitive detection and amplification of individual DNA sequences. To generate specific probes for genes or cDNAs that have not yet been cloned, it is often necessary to label PCR products which are then used in Southern or Northern hybridizations or for screening cDNA and genomic DNA libraries. In this paper a rapid and versatile method of using PCR products, as specific probes, is described, after digestion with EcoRI in buffer H, in the presence of PCR reaction buffer, and purification of the PCR products for avoid the interference by competition of unlabelled dCTP in the directionally random labelling.
Liras
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Istituto Nazionale Neurologico C. Besta, Milan, Italy.
D001483:Base Sequence; D015342:DNA Probes; D003845:Deoxycytosine Nucleotides; D015246:Deoxyribonuclease EcoRI; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D015203:Reproducibility of Results; D013997:Time Factors
D015342:DNA Probes; D003845:Deoxycytosine Nucleotides; C024107:2'-deoxycytidine 5'-triphosphate; D015246:Deoxyribonuclease EcoRI