Recombinant DNA Technology

Recombinant DNA Technology

the amplification, sequencing, and modification of a specific DNA piece

What process would you use to separate DNA fragments?

gel electrophoresis

Renaturation/Hybridization

when you cool the solution, DNA strands will bind back together

Southern Blotting

probe for a DNA fragment

Northern Blotting

probe for an RNA fragment

Western Blotting

probe fro a protein

DNA insertion into plasmid

DNA is covalently linked by ligase in the circular ds plasmid DNA

Clones

many copies of purified plasmids isolated from lysed bacteria that contain introns (recombinant plasmids)

What do clones give rise to?

DNA libraries

Genomic DNA

DNA with introns that will not provide much info on the coding region

cDNA Libraries

contain only mRNA sequences whcih is isolated and converted to dsDNA using RT

Which has information only about genes that are expressed in that cell at that time?: genomic DNA or cDNA?

cDNA

1 multiple choice option

Ingredients for DNA Synthesis in PCR

template DNA, nucleoside triphosphates, primers, and taq

PCR Steps

denature DNA by temp increase, allow primers to anneal by lowering the temp, and synthesis DNA by raising the temp again

DNA Sequencing

requires a special nucleotide without 3' hydroxyl (ddNTP)