37, No. Still, in one experimental step, the recombinants among a number of randomly selected transformants can be selected and grown, by inoculation of small liquid cultures in two 24-wells plates. Figure 3: Blue-white color selection of recombinant bacteria using X-gal. 6, pp. The selection for plasmid transformants utilizes their ampicillin resistance whereas selection for recombinant plasmids is based on the inactivation of the rpsL gene contained on the plasmid. The lacZ gene in the vector may sometimes be non-functional and may not produce β-galactosidase. TRANSFORMATION: Process of introduction of cloning vector into the host cell. Selection and Screening of Transformants. Routinely a first PCR screen of the transformants is based on the detection of stable integration of the selection marker gene. Manjula R (2004) Positive selection system for identification of recombinants using α- complementation plasmids BioTechniques ,Vol. Recombinant DNA Technology / By top. … Selection of recombinants due to inactivation of antibiotics is a laborious process as it requires:-(i) a vector with two antibiotic resistance marker (ii) preparation of two kinds of media plate, with one antibiotic each. Select the recombinant cells in the white colonies to culture. A vector is chosen where … Limitations of blue-white screening . Akamatsu T, Sekiguchi J. Analysis of transformants 1 Overview After the regeneration of protoplasts and two rounds of selection on antibiotic-containing medium the transformants can be analyzed for stable integration of the transgene by PCR-based methods. Both (ampicillin medium containing) plates are inoculated with the same transformants, but one of the two plates contains additionally a second antibiotic: tetracyclin. Selection methods in bacilli for recombinants and transformants of intra- and interspecific fused protoplasts. A transformant is a cell that has taken up additional DNA, usually a plasmid that confers some kind of antibiotic resistance. Selection of the transformed cells from the non-transformed population is done by using selectable marker genes. Selection by Antibiotic Resistance Genes. Transformant cell is that cell in which transformation has occured. The blue-white technique is only a screening procedure; it is not a selection technique.

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