General DNA manipulations were performed using standard procedures. Plasmid constructs used in this study (Table S8 ) were generated by cloning PCR products (Kapa Hifi Polymerase, Roche) obtained with oligonucleotide primers listed in Table S8 . Detection probes for SaPI DNA in Southern blots were generated by PCR using a non-proofreading polymerase (DreamTaq polymerase, ThermoFisher) using oligonucleotides specified in Table S9 . Probe labelling and DNA hybridization were performed following the protocol provided with the PCR-DIG DNA-labelling and chemiluminescent detection kit (Roche).
Pcr dig dna labelling and chemiluminescent detection kit
Manufactured by Roche
The PCR-DIG DNA-labelling and chemiluminescent detection kit is a laboratory tool used for the detection and analysis of DNA sequences. It provides a method for labeling DNA with digoxigenin (DIG) and subsequent detection using a chemiluminescent system.
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4 protocols using pcr dig dna labelling and chemiluminescent detection kit
1
Detailed DNA Manipulation Protocols
2
Time-course Phage DNA Detection
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Strains lysogenic for phages wild-type 80α and Δ(Pith-1+ith-1) were mitomycin C induced (1 μg ml−1), and 1 ml of each culture at different time points after induction was collected and used to prepare standard mini lysates, which were resolved on a 0.7% agarose gel, southern blotted and probed for phage DNA. 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). Primers are listed in Supplementary Data 1e .
3
Standard DNA Manipulation Procedures
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General DNA manipulations were performed using standard procedures. The oligonucleotides used in this study are listed in Supplementary Table S2 . 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).
4
Plasmid Construction and Southern Blot
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Plasmid constructs used in this study (Table S3 ) were generated by cloning PCR products (Kapa Hifi Polymerase, Roche) obtained with oligonucleotide primers listed in Table S4 and digested with the indicated restriction enzymes (New England Biolabs). Detection probes for phage DNA in Southern blots were generated by PCR using a non-proofreading polymerase (DreamTaq polymerase, ThermoFisher) using oligonucleotides specified in Table S4 . Probe labelling and DNA hybridisation were performed following the protocol provided with the PCR-DIG DNA-labelling and chemiluminescent detection kit (Roche, catalogue number 11093657910).
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