DNA is put into a vector - usually plasmids or a bacteriophage - and inserted and propagated in an organism. First the DNA is cut up with restriction enzymes. Then the DNA fragments are joined together in a vector. Finally, the DNA and vector are introduced into a cell via DNA transformation

Sticky ends result. These permit circularization of the DNA restriction fragment by complimentary base pairing (since each sticky end should be complimentary to the other)

Blunt ends form. These ends don't exhibit any kind of stickiness and real ability to bind so they're actually relatively stable

A vector is a DNA molecule into which another DNA fragment can be cloned; it is a carrier for recombinant DNA

As the cell replicates its own DNA the recombinant molecule is also replicated. Also, as the cell divides the recombinant molecule is transmitted to the progeny cells. A DNA fragment is said to be cloned once a transformant (the product that has been changed as a result of the added DNA) containing the recombinant molecule has been isolated

1. It can be introduced into a host cell relatively easily. 2. It contains a replication origin and can therefore replicate inside the host cell. 3. It contains a mutliple cloning site or polylinker with unique cleavage sites for many different restriction enzymes that enables many types of restriction fragments to be inserted. 4. Cells containing the vector can usually be selected by a straightforward assay, most easily by allowing growth of host cell on a solid selective medium

Plasmids are best for small DNA fragments, bacteriophages are better for larger segments and cosmids are most useful for some of the largest segments that exist

The LacZ gene is disrupted and the beta-galactosidase is made improperly

X-gal will be cleaved by B-gal and turn blue when grown in the lab

BACs are Bacterial artificial chromosomes. They're specialized vectors that can carry very large DNA fragments (which are collectively known as simply artificial chromosomes). They need to have more than 300 kilobases in order to be maintained

YACs are yeast artificial chromosomes. They incorporate essential features of linear chromosomes and include several genetic elements including a cloning site, a yeast centromere and replication origins and genetic markers which are each selectable in yeast or E. Coli. YACs up to 1MB can be recovered. YACs and BACs are essentially the same thing but YACs need many more base pairs in order to be recovered

The mapped clones method used genome "landmarks" which allowed organization which sequenced data into chromosomes. The genome was fragmented into large pieces of more than 100KB and put into YACs or BACs. BACs and YACs were then aligned to the map by sequencing short bits. Once the pieces were aligned the BACs and YACs were fragmented and put into plasmids for sequencing about 1000 bases at a time

DNA was fragmented into small pieces and each small piece was immediately sequenced. A computer was then used to find all of the overlapping fragments

There are about 20,000-25,000 protein coding genes but they make up only 2% of the entire genome. Repeat sequences make up at least 50% and 99.9% of coding sites are identical across all humans

It can help to understand the genetic basis of diseases and it can also help to identify a person's individual potential for diseases which could result in personally-tailored pharmaceuticals

There is the privacy and confidentiality issue (could this info be used by insurers, employers, courts, schools, adoption agencies, military, etc), people's reproductive decision making may be altered and the efficiency of the genetic associations could be questioned

Comparing the human genome to other primates reveals what makes us unique, small genomes help us understand what non-coding DNA is for, model organism genomes accelerate basic and applied research and sequences of micro-organisms uncover biodiversity, identify new enzymes and find new antibiotics

Functional genomics focuses on genome-wide patterns of gene expression or biochemical interactions between large numbers of genes. It tries to understand how gene expression of multiple genes is coordinated

A DNA microarray is a system of monitoring more than 10,000 levels of individual gene expression at the same time. Each spot represents a different immobilized DNA sequence (or probe)

DNA microarrays can be used to detect things like healthy cells vs diseased cells amongst other factors. Typically RNA is extracted from any two types of cells or treatment groups and the gene expression profiles of the two groups are compared

cDNA is a copy of mRNA

Sample 1 is labeled as green and sample 2 is labeled as red. If a spot is green then there is lots of sample 1. If the spot is yellow then there is a mix of expression and if the spot is red then there is overexpression of sample 2 in comparison to sample 1

A Y2H assay is a yeast two hybrid assay. It's used for identifying interacting proteins and for identifying the functions of unknown proteins since through this method you can find out which other proteins your target protein interacts with

Two-hybrid screening is used to discover protein-protein interactions. It requires the activation of a downstream reporter gene by the binding of a transcription factor to an upstream activating sequence (UAS). Both a bait protein and a prey protein must be active in this case for this sort of interaction to take place with only the bait or the prey the transcription will not take place

Once many pairs of proteins have been identified by Y2H and other methods they can be linked together to form networks. Network analysis allows proteins to be grouped by probable common function