Genomics I

Example of a cloning strategy

extract genomic DNA or mRNA

cut genomic DNA with restriction enzymes

cut vector DNA with the same restriction enzymes

ligate genomic DNA into vector (library)

transform ligated DNA into bacterial host (library)

screen for DNA inserts

select and test for a specific clone

The library

Ge

cell-based DNA cloning (in vivo)

1) cutting a target DNA

2)inserting the target DNA into the plasmid

3) introducing the plasmid to a bacteria/yeast

4) identifying the bacteria/yeast containing the plasmid with the insert

cell-free DNA cloning

in vitro method to allow selective cloning of a specific target DNA sequence wihin a heterologous collection of genomic DNA sequences

polymerase chain reaction (PCR) to amplify a specific gene or target sequence

need to design PCR primers to amplify the target sequence