Replica-plating

Shiaris MP, JJ Cooney (1983) Replica plating method for estimating phenanthrene-utilizing and phenan-threne-cometabolizing microorganisms. Appl Environ Microbiol 45 706-710. 

Figure 13.18 Replica plating for the selection of bacteria containing a recombinant vector, (a) The method used to transfer colonies of bacteria between plates, (b) Comparison of the distribution of the colonies between the plates enables identification of the colonies containing the recombinant vector. The dotted circles represent colonies that do not grow on tetracycline.

After these have produced small colonies, a replica plate is made on a tetracycline-containing medium. 

Another procedure for amplification cuts DNA containing the segment of interest into small pieces with a restriction enzyme. The cut pieces are incorporated into a plasmid or virus vector to be amplified in a suitable host. After growth, the mixture is plated to produce a mixture of bacterial or viral clones. The clone or clones of interest are identified often by hybridization of the clones after replica plating with a radioactive probe, followed by autoradiography to find the clone of interest. 

Replica plating. A technique in which an impression of a culture is taken from a master plate and transferred to a fresh plate. The impression can be of bacterial clones or phage plaques. 

Incubate the cultures at 37°C with shaking overnight. These strains will be used on Day 3 to isolate plasmid DNA. Place the two replica plates produced in step 3, agar side up, in the 37°C incubator overnight. 

Figure 2.6. Replica plating and in situ hybridization to identify colonies that contain the DNA of interest.

Figure 4 The 384-channel parallel multidispensing of (a) 1 jiL and (b) 200 nL aliquots out of one aspiration into empty wells. The false color maps on the left show the DFM values of the dispensed aliquots over the plate. The color code for the DFM values is shown in a look up table (vertical bars on the right-hand side of the two plate maps). The histograms show the distribution of the DFM values around the mean (average) aliquot volume. The false color map, the DFM histogram, and the stated values for %Bias, %CV, and min/max %DFM in (a) and (b) refer to one representative plate from the 24 replica plates produced in a replication cycle for (a) 1 pL and (b) 200 nL aliquots, respectively. Roche uses the first 2 columns (2 X 16=32 wells) in each 384-well plate for standards and controls since columns 1 and 2 are not filled with compounds, they are not included in the false color maps...

Transfer colonies to a replica plate Lyse bacteria to expose proteins... 

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