Construction of genomic library using lamda phage
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11 سال پیش
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A genomic library is a collection of the total genomic DNA from a single organism. The DNA is stored in a population of identical vectors, each containing a different insert of DNA. In order to construct a genomic library, the organism's DNA is extracted from cells and then digested with a restriction enzyme to cut the DNA into fragments of a specific size. The fragments are then inserted into the vector using the enzyme, DNA ligase.[1] Next, the vector DNA can be taken up by a host organism- commonly a population of Escherichia coli or yeast- with each cell containing only one vector molecule. Using a host cell to carry the vector allows for easy amplification and retrieval of specific clones from the library for analysis.[2]
There are several kinds of vectors available with various insert capacities. Generally, libraries made from organisms with larger genomes require vectors featuring larger inserts, thereby fewer vector molecules are needed to make the library. Researchers can choose a vector also considering the ideal insert size to find a desired number of clones necessary for full genome coverage.[3]
Genomic libraries are commonly used for sequencing applications. They have played an important role in the whole genome sequencing of several organisms, including the human genome and several model organisms.[4] Genomic libraries are also utilized in comparison studies between differing species.
Phage λ is a double-stranded DNA virus that infects E. coli. The λ chromosome is 48.5kb long and can carry inserts up to 25kb. These inserts replace non-essential viral sequences in the λ chromosome, while the genes required for formation of viral particles and infection remain intact. The insert DNA is replicated with the viral DNA; thus, together they are packaged into viral particles. These particles are very efficient at infection and multiplication leading to a higher production of the recombinant λ chromosomes.[3] However, due to the smaller insert size, libraries made with λ phage may require many clones for full genome coverage. Source of the article published in description is Wikipedia. I am sharing their material. Copyright by original content developers of Wikipedia.
Link- http://en.wikipedia.org/wiki/Main_Page
http://shomusbiology.weebly.com/
Download the study materials here-
http://shomusbiology.weebly.com/bio-m...
A genomic library is a collection of the total genomic DNA from a single organism. The DNA is stored in a population of identical vectors, each containing a different insert of DNA. In order to construct a genomic library, the organism's DNA is extracted from cells and then digested with a restriction enzyme to cut the DNA into fragments of a specific size. The fragments are then inserted into the vector using the enzyme, DNA ligase.[1] Next, the vector DNA can be taken up by a host organism- commonly a population of Escherichia coli or yeast- with each cell containing only one vector molecule. Using a host cell to carry the vector allows for easy amplification and retrieval of specific clones from the library for analysis.[2]
There are several kinds of vectors available with various insert capacities. Generally, libraries made from organisms with larger genomes require vectors featuring larger inserts, thereby fewer vector molecules are needed to make the library. Researchers can choose a vector also considering the ideal insert size to find a desired number of clones necessary for full genome coverage.[3]
Genomic libraries are commonly used for sequencing applications. They have played an important role in the whole genome sequencing of several organisms, including the human genome and several model organisms.[4] Genomic libraries are also utilized in comparison studies between differing species.
Phage λ is a double-stranded DNA virus that infects E. coli. The λ chromosome is 48.5kb long and can carry inserts up to 25kb. These inserts replace non-essential viral sequences in the λ chromosome, while the genes required for formation of viral particles and infection remain intact. The insert DNA is replicated with the viral DNA; thus, together they are packaged into viral particles. These particles are very efficient at infection and multiplication leading to a higher production of the recombinant λ chromosomes.[3] However, due to the smaller insert size, libraries made with λ phage may require many clones for full genome coverage. Source of the article published in description is Wikipedia. I am sharing their material. Copyright by original content developers of Wikipedia.
Link- http://en.wikipedia.org/wiki/Main_Page
11 سال پیش
در تاریخ 1392/08/03 منتشر شده
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