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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 blue-white technique is only a screening procedure; it is not a selection technique. Transformant cell is that cell in which transformation has occured. Limitations of blue-white screening . Manjula R (2004) Positive selection system for identification of recombinants using α- complementation plasmids BioTechniques ,Vol. 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. 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. … Selection by Antibiotic Resistance Genes. The lacZ gene in the vector may sometimes be non-functional and may not produce β-galactosidase. Selection of the transformed cells from the non-transformed population is done by using selectable marker genes. TRANSFORMATION: Process of introduction of cloning vector into the host cell. 6, pp. Akamatsu T, Sekiguchi J. Figure 3: Blue-white color selection of recombinant bacteria using X-gal. 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. Selection methods in bacilli for recombinants and transformants of intra- and interspecific fused protoplasts. 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. Select the recombinant cells in the white colonies to culture. 37, No. A transformant is a cell that has taken up additional DNA, usually a plasmid that confers some kind of antibiotic resistance. Recombinant DNA Technology / By top. Both (ampicillin medium containing) plates are inoculated with the same transformants, but one of the two plates contains additionally a second antibiotic: tetracyclin. A vector is chosen where …

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