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A Complete Guide to Gene Cloning: From Basic to Advanced Selection, Screening, and Analysis of Recombinant Clones

A Complete Guide to Gene Cloning: From Basic to Advanced : Selection, Screening, and Analysis of... [In this chapter, an attempt has been made to discuss the most commonly used screening and selection techniques of recombinants. Once the gene is transferred to the host cell, the challenge lies in whether the gene of interest is present or not, and if it is present, it can express itself or not. Selection can be defined as creating an environment where only transformants and more specifically the recombinants survive. It also involves choosing the selected hosts for the target gene which express in the defined environment as well as through product formation mainly through protein. The techniques involved in the selection and screening of recombinants that are practiced in the laboratory are explained with a diagrammatic illustration. Selection and screening cannot be generalized. Each method has its own advantages and limitations. The success of genetic engineering experiments depends on the expression of the gene of interest in the host cells. Also, the chapter highlights high throughput screening techniques which have scope for automation and analysis at a faster rate. By the end of the chapter, the methods to characterize the identified clones for further downstream activities are discussed.] http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png

A Complete Guide to Gene Cloning: From Basic to Advanced Selection, Screening, and Analysis of Recombinant Clones

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Publisher
Springer International Publishing
Copyright
© The Editor(s) (if applicable) and The Author(s), under exclusive license to Springer Nature Switzerland AG 2022
ISBN
978-3-030-96850-2
Pages
97 –117
DOI
10.1007/978-3-030-96851-9_7
Publisher site
See Chapter on Publisher Site

Abstract

[In this chapter, an attempt has been made to discuss the most commonly used screening and selection techniques of recombinants. Once the gene is transferred to the host cell, the challenge lies in whether the gene of interest is present or not, and if it is present, it can express itself or not. Selection can be defined as creating an environment where only transformants and more specifically the recombinants survive. It also involves choosing the selected hosts for the target gene which express in the defined environment as well as through product formation mainly through protein. The techniques involved in the selection and screening of recombinants that are practiced in the laboratory are explained with a diagrammatic illustration. Selection and screening cannot be generalized. Each method has its own advantages and limitations. The success of genetic engineering experiments depends on the expression of the gene of interest in the host cells. Also, the chapter highlights high throughput screening techniques which have scope for automation and analysis at a faster rate. By the end of the chapter, the methods to characterize the identified clones for further downstream activities are discussed.]

Published: Apr 27, 2022

Keywords: Nucleic acid hybridization; Immunological detection; Restriction mapping; Positive and Negative selection; DNA sequencing

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