PCR and AGE

tools of molecular biology

-Process of making multiple copies of
target DNA sequence
-- is closest to DNA replication cuz it creates copies of specific DNA sequences
-in should know the DNA sequence you're targeting


-in vitro replication (cell free or outside the cell)

PCR helps amplify the target
segment, making millions of DNA
copies in 1 hour

what part of central dogma is closest to pcr

3 steps of pcr

source for PCR. Only one enzyme or DNA polymerase

95°C

temperature will lower to 55-60°C primers will attach to separated DNA strands. The primers are the series of sequences (usually 8 to 10 bases) and it will attach to the DNA template. Cuz DNA polymerase can't stand the temp

Temp will increase to 72°C where complementary bases will attach to each other.
Dna samples (the extracted saliva)

Products of PCR are called. what u detect using AGE or agarose gel electrophoresis

Ampicons or Amplification products are detected using

the equation for cycles in the PCR

Determine the length of the DNA strands/fragments

The gel in AGE is made from agar à acts as a filter to
sort DNA strand by size
-> Porous in nature

Electric current is applied to the gel to make the DNA
migrate/move across the gel
à Negative to positive charge

Shorter DNA = “moves” faster
Longer DNA = “moves” slower

The gel in AGE is made from ?
-> acts as a filter to sort DNA strand by size
-> Porous in nature

Electric current is applied to the gel to make the DNA
migrate/move across the gel in AGE
?

Shorter DNA = “moves” ?
Longer DNA = “moves” ?

Agarose gel electrophoresis determines the length of DNA strands from the negative to the positive side. This is because DNA is ? because of the structure of the nucleotides.

is the standard for u to know the size of your DNA ( 5bp (base pairs) 100 BP, 1000 BP, 10000 kb.

Use these ? BP if u know your DNA is small.

use this if u have a lot of base pairs and if DNA fragment are large (high molecular weight markers)

bp and kp (base pair). e should be able to tell the size of the DNA sample from the ?

Relationship between percent relationship of agarose (size of pores) to DNA sizes is ?.

The higher the percentage of the agarose, the smaller the pores and DNA fragments u can run thru the gel. And opposite. Not small cuz the small DNA will go thru the gel and u won't be able to observe them.

Differently sized molecules will occupy different parts of ? from largest at the top to smalled at the bottom.

eukaryotes is in ? in the cell

prokaryotes are in? in the cell

What is used to better know the quantify and quality of the DNA extracted