1
Q
Which technique is required to carry out a RFLP assay?
A
Southern blot
2
Q
southern blots use what type of probes?
A
nucleic acid probes.
3
Q
PCR analysis
A
- Taq polymerase to copy DNA b/w primers.
- Remelt, repeat reaction multiple times (amplification)
- It is highly specific
4
Q
DNA Sequencing
A
- Info on specific nucleotides
- Takes a long time, expensive
5
Q
RNA Methods: Northern Blot
A
- Denature RNA to make it linear
- Nucleic acid probe complementary to RNA
- Unlike southern blot, you can QUANTIFY the amount (of RNA though, not DNA)
6
Q
RNA Methods: RT- PCR
A
- Converts mRNA into DS DNA using reverse transcriptase and a primer
- Exonuclease degrades old mRNA
- Run DNA in PCR assay.
7
Q
RNA Methods: Microarray
A
- Labeled RNA hybridized to unlabeled probe (opposite of northern blot)
- mRNA with probe then hybridized on a spot-plate.
- Idea of HOW MUCH mRNA is present.
- Large-scale analysis
8
Q
Southern vs. Western vs. Northern: Differences in what they do?
A
- Western blot uses antibody probe, whereas southern and northern use nucleic acid probes
- Northern and Western use denaturing gels, whereas southern does not
- Southern (RFLP analysis) used for presence of DNA sequence, NOT amount of DNA
- Northern can quantify amount of RNA, and has to make RNA linear (denaturing)
