Analyzing Cells, Molecules And Systems 2 Flashcards
HindIII Characteristics
- Creates 4 “Sticky” ends
- Cleaves AA sequence in the AAGCTT sequence
5’-3’
Purpose of DNA Microassays?
Used to determine the overall change in gene expression b/w 2 samples
Cool thing about DNA Libraries
- Every gene in the human body can be put into bacteria
- Can be stored
- Can be INDEFINITELY propagated at anytime
EcoRI Characteristics
- Creates 4 “sticky” ends
- Cleaves before the AATT sequence
5’-3’
Purpose in Detection variances in DNA sequences
- Forensics
- Diagnostics
- Pre-Natal
- Newborn screening
- Genetic carriers
PCR Products at the end of the
- 1st cycle
- 2nd cycle
- 3rd cycle
- 2
- 4
- 8
The main Restriction Endonucleases
- HaeIII
- EcoRI
- HindIII
Genomic Library vs cDNA Library
Genomic:
- Contains introns
- produces DNA fragments
cDNA:
- No introns
- Much smaller than original gene
- Req Reverse transcriptase
Purpose of PCR
- Amplify Isolated DNA regions
- Need Primers
- Heat to anneal then cool to replicate
- Earlier detection of microorganisms
(HIV, bacterial and fungal infection) - Detection of Specific genetic mutations
T/F: Genes can be cloned using Bacteria.
True
- W/ the use of PLASMIDS
RFLP is also called what?
- DNA finger printing
- DNA typing
HaeIII Characteristics
- Cleaves straight Down at a GGCC site
5’-3’
Why is Short Tandem Repeats (STRs) interesting?
Most individuals have VARYING repetitive regions of DNA sequences
Ligaments Rxn req. ___ to work.
ATP
What is Quantitative PCR?
Used to Quantify copy number of a specific gene in 2+ samples in real time
- Includes a PROBE which fluorescent only in the presence of the PCT Product
Diff b/w Gel Electrophoresis and SDS Page
Gel:
- Analyzes DNA
- DNA already Negative
SDS:
- Separates Proteins
- Needs to Negatively charge them first
What is Quantitative PCR used for?
- Detect Levels of infectious agent
- Determine levels of gene expression
How can you glue 2 fragments that were cus w/ diff restriction nucleases?
- DNA Poly + dNTPs
- Ligase + ATP
Restriction Endonuclease
- Enzymes isolated from Bacteria
- Cut DNA at Specific sites
3 detections of variations in DNA sequences
- Restriction Fragment Length Polymorphism
(RFLP) - Variable Number of Tandem Repeats (VNTR)
- Short Tandem Repeats (STR)
How can HIV be diagnosed?
PCR
