Genomic Libraries use what to (4) clone DNA?

1. Plasmids (10K bp)

2. Bacteriophages (25K - 100K bp)

3. Cosmids (50K bp)

4. Bacterial Artificial Chromosomes (100K - 1 mill. bp)

Cosmid

bacteriophage sequence called Cos sites that act as origins of replication clones.

What are the 3 steps to a Genomic Library?

1. Cleave DNA from human with restriction nuclease

2. Insert fragments into plasmids via ligase

3. Introduce Plasmid into Bacteria

What do Genomic Libraries allow us study in Eukaryotes (3)?

1. intergenic regions

2. sequences involved in controlling gene expression

3. introns of the gene

What do cDNA libraries allow us to determine in (3) Eukaryotes?

1. exons of the gene

2. deduce the protein sequence using mRNA

3. mutation/expression of eukaryotic genes in prokaryotic cells.

What is Southern Blotting used for?

DNA targeting by a DNA, cDNA or RNA probe

What is Northern Blotting used for?

RNA targeting by a DNA, cDNA or RNA probe.

What is Western blotting used for?

Protein targeting by an antibody

What is Restriction Mapping used for?

Digest DNA and separate bands

What is Restriction Fragment Length Polymorphisms (RFLP) used for?

genetic mapping of disease alleles and for DNA fingerprinting.

What is Variable Number Tandem Repeats (VNTR) used for?

paternity and forensic analyses; analyzes loci that have variable numbers of tandem repeats.

What are Microsatellites?

tandem repeats about 2-5 bp's long. Present in hundreds of tandemly arrayed copies.

What are Microsatellites used for?

to perform analysis for trinucleotide repeat disorders.

What are Single Nucleotide Polymorphisms (SNP) used for?

mapping genetic disorders that might be linked to a specific SNP.

What is functional genomics?

strategies that are designed to determine the function of individual genes.

Transcriptomics

looks at gene expression @ the mRNA level.

(mRNA increases in a certain gene..maybe that gene plays a role)

Proteomics

looks @ gene expression at the protein level.

knows what proteins are expressed when.

What are the areas of Functional Genomics (3) ?

1. Bioinformatics

2. Mutagensis

3. Protein-Protein Interactions

Bioinformatics

Takes proteins from 1 organism and compares it to another via supercomputer.

Views protein forms.

Protein-Protein Interactions

Removes the domain of proteins that cause interaction and replaces it w/more proteins and sees if the proteins will then interact.

Mutagenesis

Mutate the gene found to test for virulence or other issue and mutate it again to see how it performs.

Microarray Anaylsis

2D protein electrophoresis

What is Signature Tag Mutagenesis used for?

to figure out which genes are involved in virulence (pathogens) 

High Throughput Analysis - Bulk