dna hybridization

hybridization

double stranded formation or annealing between two different single strands of DNA or RNA, can also be RNA and DNA due to complementary base pairing

plotting technique

it detects specific biomolecules like nucleic acids as well as proteins in complex samples

probes

these are often used to study nucleic acids and other important biomolecules

DNA or RNA probe

short segments of nucleic acid of known base sequences

southern blot

enables efficient analysis of DNA fragments in an immobilized membrane support such as nitrocellulose membrane

Edwin M. Southern

he first described the southern blot technique of DNA analysis 1975

true

t/f: gel electrophoresis sorts fragments according to their size

alkaline solution

it is used to separate the double strands in southern blotting

saline sodium citrate (ssc buffer)

buffer used for the blotting technique

capillary action and ion exchange interaction

driving forces to transfer the DNA from the gel to the membrane

70 to 80 C for 2 hours

to immobilize the nucleic acid, it should be incubated for?

true

t/f: during the immobilization step, uv radiation may be used for nylon membrane only

buffer

after hybridization, excess probe is washed from the membrane by?

x-ray

used to visualize a radioactive material

uv light

used to visualize fluorescent material

northern blot

used to detect the RNA of interest and study gene expression

mRNA

usually detected by northern blot

false

t/f: DNA replication is a process of gene expression

true

t/f: either RNA or DNA probe may used for northern blotting as long as it is complementary to the target RNA

James Alwine, David Kemp, and George Stank

developed northern blot in 1977

oligo deoxythymine

hybridizes the poly A tail of the mRNA to isolate it

formaldehyde

used for denaturing step in northern blot

formamide

lowers annealing temperature, eliminating the need for high temperatures which could cause RNA degradation

nylon membrane

membrane used for northern blot