Scribd
Upload
104 views

Module IV - Genomic DNA and cDNA Library

A genomic DNA library is a collection of DNA fragments from an organism's entire genome cloned into a host. Construction involves isolating genomic DNA, fragmenting it, ligating the fragments into a vector, and transforming the vector into a host. This allows researchers to explore genomic structure/function, map genes, and identify mutations or pharmaceutical genes. While powerful, genomic libraries can be error-prone for large eukaryotic genomes containing non-coding DNA.

Uploaded by

Ananya Singh
Copyright
© © All Rights Reserved
Available Formats
Download as DOCX, PDF, TXT or read online on Scribd
104 views

Module IV - Genomic DNA and cDNA Library

A genomic DNA library is a collection of DNA fragments from an organism's entire genome cloned into a host. Construction involves isolating genomic DNA, fragmenting it, ligating the fragments into a vector, and transforming the vector into a host. This allows researchers to explore genomic structure/function, map genes, and identify mutations or pharmaceutical genes. While powerful, genomic libraries can be error-prone for large eukaryotic genomes containing non-coding DNA.

Uploaded by

Ananya Singh
Copyright
© © All Rights Reserved
Available Formats
Download as DOCX, PDF, TXT or read online on Scribd
You are on page 1/ 4

MODULE IV: Genomic DNA and cDNA libraries

Genomic DNA library


The genomic DNA library is a collection of comprehensive DNA (including
introns and exons) fragments representing the entire genome of a species.
It is complete genome in multiple clones containing small DNA fragments.
Depending upon organism and size of genome, this library is either prepared in
a bacterial vector or in YAC
Construction of a genomic DNA library involves four step:
1. Isolation of genomic DNA
2. Generation of suitable size DNA fragments
3. Cloning in a suitable vector system (depending upon size)
4. Transformation in suitable host.
 A sample containing the complete genome is mixed with with
restriction enzymes (HaeIII and AluI). These enzymes cleave the large
DNA molecule into smaller gene fragments.
 The smaller gene fragments are now separated on the basis of size
(size fractionation) by electrophoresis.
 EcoR1 methylase is added to block EcoR1 sites present within the
fragments.
 This is followed by blunt end ligation with EcoR1 linker molecules,
followed by restriction by EcoR1 enzymes to convert the blunt ends
to cohesive ends.
 Anneal the cohesive ends of smaller fragments with natural cohesive
ends of λ Charon 4A DNA (replacement vector)
 T4 DNA ligase enzyme is used to ligate the chosen DNA sequence
into the vector.
 The vector is then inserted into E.coli.
 These cells then replicate their genome along with the vector
genome contained within them.
 The collection of clones that contain all sequences from original
source forms genomic library.
Advantages of a Genomic Library
 Genomic libraries derived from eukaryotic organisms are critical for
studying the genome sequence of a particular gene of interest.
 It is useful for prokaryotes with small genomes to identify a clone
encoding a specific gene of interest.
 It helps researchers to explore more about an organism’s genomic
structure and function. It is also used to study genetic mutations.
 Pharmaceutically important genes can also be identified by this
method.

Disadvantages of a Genomic Library


 It requires sophisticated software and a vast amount of computing
power. Also, the process is prone to errors.
 Eukaryotic genome libraries with very large genomes contain many
DNA that do not code for proteins, as well as non-coding DNA like
repetitive DNA and regulatory regions, making them less than ideal.

Uses of Genomic Library


 Researchers can explore the genome of an organism to learn more
about genomic structure and function
 They can map the genome, identifying the locations of specific genes.
 Helps to develop tests which can be used to locate genetic variations
including mutations
 Useful in Recombinant DNA Technology, helps to genetically modify
organisms and produce clones of desired types.
 Genomic library construction is the first step in any DNA sequencing
projects.
 Genomic library helps in identification of the novel pharmaceutically
important genes

You might also like