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Terms in this set (25)
Probe: Sequence of ___ in which each molecule has been labeled with radioactive ____.
Southern Blot: ___ cleavage, electrophoresis, transfer to nitrocellulose, and ___ with a ____
DNA fragments separated in a gel can be
transferred to a ____ for _____ to a SS DNA probe
The extent of hybridization can be quantified by using a radioactive ____ ____ and auto-radiography
what is the advantage to using a southern blot?
can hybridize DNA with small amount of DNA
Nucleic Acid Hybridization ___ ___ allow fragment
to be separated from the others and purified...
Can be done by ____, but it's a time consuming work
If the fragment of interest is not too long, it's possible to obtain large quantities of the fragment by ____
KEY features of DNA replication:
An enzyme that forms the sugar-phosphate bond is called a ___ ____
1.Can only _____, can not initiate synthesis ofnew strand: needs _____!
2. Only add nucleotides to ____end
DNA synthesis requires ___ _____ & ____
PCR requires _____ primers, one at ___ ___
Each cycle ___ the amount of target DNA
Repeated cycles produce millions or billions of copies
Each PCR cycle
has 3 steps-
a.Melting of DNA
(____ 95 C)
b. Hybridization of primer
(_____: 50 to 60 C)
c. DNA synthesis
(______: 70 C)
DNA Polymerases from Archaea: _____ is commonly used.
-Requires only ____ amounts of ___.
-___ and reliable - criminal investigation/blood bank
-____ is much higher than nucleic acid hybridization
- Sequences at the ends of region must be ____
- Sequences longer than 5000 bp are ___ ___ ___
not efficiently amplified
- Genotype- The ___ ___ of an organism.
- Phenotype- The ___ ____ of an organism.
-Homologous chromosomes- in a ____ organism, the 2 copies
of a chromosome inherited from the ____ and the ____.
- Locus- ___ of a gene on a ____.
- Allelomorph (allele)- ___ versions of the same _____.
ex) GCA - VALINE | GCG - VALINE
- Homozygous- the 2 copies of a gene are ____.
- Heterozygous- the 2 copies of a gene are ____.
Genetic variations exists in most natural populations
of organisms... because multiple ____ exists for many _____.
We called such genetic differences as DNA _____ or genetic markers or DNA _____.
Types of DNA markers present in genomic DNA:
- Single nucleotide polymorphism (SNP)
- Restriction fragment length polymorphism (RFLP)
- Amplified Fragment length Polymorphism (AFLP)
- variable number of tandem repeats (VNTR)
READ READ READ THROUGH
•SNPs are single-nucleotide polymorphisms or ___ ____variations in the DNA sequence
These are ___ ____ form of differences among
people.....we know some 3 million SNPs
In human genome, there is a SNP for every ______bp in NON-coding region, one every ______ bp in CODING region.
what tool is used to identify SNPs
Most SNPs require DNA _____ to study, unless it's located in the ______ site
A SNP that eliminates a restriction site is called " ____"
- Restriction Fragment Length Polymorphism
•Differences in DNA sequence on homologous chromosomes that result in restriction fragments of varying _____ that can be detected using DNA ____ (____ Blotting)
Also distinguish homozygous from ____!!
RFLP alleles are ____
RFLPs create DNA fragments of different ____.
The fragments are the same size in a ____.
The fragments are different sizes in a ____
Simple Tandem Repeat Polymorphism (STRP)
____of the entire repeated target by ____, using ____ primers that hybridize in flanking DNA. More repeats lead to larger PCR product.
Randomly amplified polymorphic DNA (RAPD)
Most methods require prior Knowledge of The ___
RAPD are ____ markers
RAPD: Randomly amplified polymorphic DNA
Cheap, fast, __ ___ ___ of the
sequence is required.
- Too ____......may not produce the same Band
- Sometimes produce too ___ bands
NO prior knowledge
AFLP: Amplified Fragment length Polymorphism
- ____ amplification
More reliable and relatively evenly distributed
Produce ____ number of bands
- Little more expensive
- Requires use of ___ ___
Copy Number Polymorphism (CNP's)
A normal variation in DNA due to variation in the number of copies of a sequence within the DNA
(extra or missing copies of the genome of 1 kb to 1Mb)
Typically individual only have 2 copies!!
CNP's associated with hemoglobin genes....associated
With resistance to _____
CNP's with HIV receptor gene CCL3 associated with
Resistance to _____
Disease genes can be mapped using __ ___
Therefore, it is easier to map a new gene using a DNA marker
applications for DNA markers
- genetic mapping and detection of disease gene
- DNA finger printing
- population studies- level of genetic variation
- improvement of domestic plants and animals
- evolutionary genetics, ecological indicators & evolutionary relationship
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