DNA Fragmentation and Cloning Vectors

4 cutters

6 cutters

8 cutters

The ends they produce don't last long

They can cut more than one sequence

Where in the sequence they cut changes

To get longer fragments and more likely preserve the gene

To save time

To prevent the DNA from degrading

Binds to pilus surface

Enters through bacterial pores

Direct injection of DNA

Is convered to dsDNA that acts like a plasmid

Is integrated into the bacterial chromosome

Is cut up by restriction enzymes ans integrated into the bacterial chromosome

It goes from blue to white in the presence of LacZ

It goes from white to blue in the presence of LacZ

It goes from white to blue in the presence of the small subunit of LacZ

The M13 phage will treat the plasmid as its own DNA and replicate it for secretion

The phage will insert the vector into the chromosome

The phage will prevent the bacteria from destroying the plasmid

Cos lambda sites

Cos beta sites

Cos gamma sites

BAC's

YAC's

MAC's

Adding AMP to ligated ends gives easily ligable high energy form

ATP & T4 remove the phosphates from the ends of the DNA to prevent self annealing

T4 can only ligate DNA in the presence of ATP