General DNA manipulations were performed using standard procedures. The plasmids and oligonucleotides used in this study are listed in S7 and S8 Tables, respectively. The labelling of the probes and DNA hybridization were performed according to the protocol supplied with the PCR-DIG DNA-labelling and Chemiluminescent Detection Kit (Roche). Southern and western blots experiments were performed by standard procedures [15 (link)].
Pcr dig dna labelling
Manufactured by Roche
The PCR-DIG DNA-labelling is a laboratory equipment used for the incorporation of digoxigenin (DIG) into DNA during the polymerase chain reaction (PCR) process. This allows for the subsequent detection and analysis of the targeted DNA sequences.
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3 protocols using pcr dig dna labelling
1
Standard DNA Manipulation Procedures
2
Standardized DNA Manipulation Techniques
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General DNA manipulations were performed using standard procedures. The oligonucleotides used in this study are listed in S4 Table . The labeling of the probes and DNA hybridization were performed per the protocol supplied with the PCR-DIG DNA-labelling and Chemiluminescent Detection Kit (Roche). Detection probes for SaPI DNA in Southern blots were generated by PCR using primers SaPIbov1-112mE and SaPIbov1-113cB (SaPIbov1 and SaPIbov5) as listed in S4 Table .
3
Standard DNA Manipulation Protocols
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General DNA manipulations were performed using standard procedures. DNA samples were heated at 75°C for 10 min prior to the electrophoresis to ensure cos site melting. The plasmids and oligonucleotides used in this study are listed in the electronic supplementary material, tables S2 and S3, respectively. The labelling of the probes and DNA hybridization were performed according to the protocol supplied with the PCR-DIG DNA-labelling and Chemiluminescent Detection Kit (Roche). To produce the phage and SaPI mutations, we used plasmid pBT2-βgal, as previously described [11 (link)].
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