Team:Gaston Day School/Experiments

Experiments and Procedures

Below are the most frequent procedures for the experiments we performed this year.

Single-temperature Double Digest

Litigation Protocol with T4 Ligase (M0202)

Plasmid Preparation Procedure

Plasmid Preparation Procedure (By Hand)

Gibson Assembly Protocol (E5510)

Modified Competent Cells Procedure

Sequential Double Digest

Transformation Procedure

SeaPhages Procedures for Isolating Environmental Phages

  • Testing the specificity of phages - T4 phage (plates in the right column of each image) grows on standard K-12 and substrain W but not on substrain C. PhiX phage (plates in the left column of each image) does not grow on K-12 or W but does grow somewhat on substrain C.

    Testing Environmental Samples for the presence of phage - Two samples (in the upper 2 sections of the plate) show cleared rings around the inoculation site which indicates that bacteriophages are likely present in the sample. The bottom section shows no cleared area, indicating a lack of bacteriophages in that sample.

    Can bacteriophages transfer plasmid DNA? The bacteria in these tubes show the presence of Red Fluorescent Protein after being infected with T4 bacteriophages grown on bacteria carrying an RFP plasmid. This shows that bacteriophages can transfer plasmid DNA just as they can transfer genomic DNA.

    Is the plasmid transferred as a plasmid or is it incorporated into the genome? Lanes 2 and 3 on the gel show the DNA isolated from transduced bacteria using a plasmid prep kit from New England Biolabs. The band in lane 4 is the plasmid isolated from the originating bacteria. This strongly supports the direct transfer of the plasmid and that the RFP and antibiotic resistance genes have not been incorporated into the genome of the bacteria.