dna libraries dr. e. what are dna libraries? a dna library is a collection of dna fragments that...

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DNA LIBRARIES Dr. E

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Page 1: DNA LIBRARIES Dr. E. What Are DNA Libraries? A DNA library is a collection of DNA fragments that have been cloned into a plasmid and the plasmid is transformed

DNA LIBRARIES

Dr. E

Page 2: DNA LIBRARIES Dr. E. What Are DNA Libraries? A DNA library is a collection of DNA fragments that have been cloned into a plasmid and the plasmid is transformed

What Are DNA Libraries?

• A DNA library is a collection of DNA fragments that have been cloned into a plasmid and the plasmid is transformed into a bacteria for propagation and storage.

• Ideally contains at least one copy of every sequence.

• Allows researchers to identify and isolate the DNA fragments that interest them for further study.

Page 3: DNA LIBRARIES Dr. E. What Are DNA Libraries? A DNA library is a collection of DNA fragments that have been cloned into a plasmid and the plasmid is transformed

What Are The Two Types Of Libraries?

• Genomic library-A set of cloned DNA fragments that collectively represent the entire genome of a given organism.

• cDNA library-represents a sample of all the expressed mRNA’s from a particular cell type, particular tissue, or an entire organism which has been converted back to DNA. Thus represents the genes that were actively being transcribed (turned on).

Page 4: DNA LIBRARIES Dr. E. What Are DNA Libraries? A DNA library is a collection of DNA fragments that have been cloned into a plasmid and the plasmid is transformed

Applications Of cDNA Libraries

• Discovery of novel genes

• Determine what mRNA’s are expressed in different cells or tissues

• Determine what mRNA’s are expressed at different developmental stages

• Study alternative splicing

Page 5: DNA LIBRARIES Dr. E. What Are DNA Libraries? A DNA library is a collection of DNA fragments that have been cloned into a plasmid and the plasmid is transformed

Applications Of Genomic Libraries

• Determine the complete genome sequence

• Study of regulatory sequences

• Study promoters

• Study intergenic regions

• Study Introns

• Study non-expressed gened

Page 6: DNA LIBRARIES Dr. E. What Are DNA Libraries? A DNA library is a collection of DNA fragments that have been cloned into a plasmid and the plasmid is transformed

Making a Genomic Library

Page 7: DNA LIBRARIES Dr. E. What Are DNA Libraries? A DNA library is a collection of DNA fragments that have been cloned into a plasmid and the plasmid is transformed

Making a cDNA Library

Page 8: DNA LIBRARIES Dr. E. What Are DNA Libraries? A DNA library is a collection of DNA fragments that have been cloned into a plasmid and the plasmid is transformed

Separating mRNA

Page 9: DNA LIBRARIES Dr. E. What Are DNA Libraries? A DNA library is a collection of DNA fragments that have been cloned into a plasmid and the plasmid is transformed

Creating cDNA

Page 10: DNA LIBRARIES Dr. E. What Are DNA Libraries? A DNA library is a collection of DNA fragments that have been cloned into a plasmid and the plasmid is transformed

Inserting cDNA Fragment into

Plasmid

Page 11: DNA LIBRARIES Dr. E. What Are DNA Libraries? A DNA library is a collection of DNA fragments that have been cloned into a plasmid and the plasmid is transformed

Where is cDNA Fragment Inserted

into Plasmid?

Page 12: DNA LIBRARIES Dr. E. What Are DNA Libraries? A DNA library is a collection of DNA fragments that have been cloned into a plasmid and the plasmid is transformed

pTriplEx Plasmid

Page 13: DNA LIBRARIES Dr. E. What Are DNA Libraries? A DNA library is a collection of DNA fragments that have been cloned into a plasmid and the plasmid is transformed

Anatomy Of A Gene