chapter 20 Flashcards
Molecular technologies (14 cards)
western blotting
measure protein size + abundance
measure protein quantity
how does western blotting work
- sample prepartion by extraction proteins from cells/tissue
- gel electrophoresis that separate proteins by size on polyacrylamide gel
- membrane blotting: transfer size separated proteins to membrane
- immunodetection:
- probe blot w. primary antibody against protein of interest
- detect primary antibody w. enzyme linked secondary antibody
Which part of a Western blotting
experiment ensures detection of
one specific protein as opposed to
a different protein?
primary antibody (a good one only binds to 1 specific target protein)
In Western blotting, the secondary
antibody binds directly to ….
primary antibody
wester blotting: smaller proteins move ____ and ___ in gel
faster and farther
- will be farthest down
PCR/polymerase chain reaction
is used to amplify detectable/ usable amounts of DNA from very small starting amounts even from a single cell
- detects viral proteins
The enzyme used in PCR is ____
DNA polymerase
3 steps of PCR cycles are
denaturation, annealing, elongation (primer extension)
In PCR, the amount of DNA ____ with each round of amplification.
doubles so
30 cycles -> 30 doublings so after 30 cycles there will be
2^30 X the number of starting template molecules = 1 billion
real time PCR
Detects the DNA amplified by PCR in real time during the PCR
measuring nucleid acid template amounts
CRISPR-Cas gene editing
cutting and editing DNA
- used for gene therapy
how CRISPR-Cas works
- Cas binds to target DNA
- Cas cleaves it at precise spots
- repair of cut DNA can give a small deletion or precise DNA change
To edit the sequence of one of a
gene’s introns using CRISPR-Cas,
the sequence of the guide RNAs
would have to be complementary to parts of ___
the intron of the gene
reverse transcriptase
amplifying RNA
