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indicator method
Presence of
Residue/Precipitate
it means there is
chemical bonding
basically produce
precipitate like
crystalline, coagulates,
granules
Solubility
use in materials that are
typically immiscible (cannot
mix each other ) or low
solubility
layer formation (lipids)
test of chemical
wherein there is a
formation of active
gas liberation
through bubble
formation called
BIOCHEMICAL TEST
identify the main important
chemical compounds
each test take a small amount of
the substance to test, shake it in
water in a test tube( the substance
may need grinding with a pestle
and mortar, to break up the cells
and release the cell contents
many of these compounds are
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BIOCHEMICAL TEST
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1.
b.
c.
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Primer
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Application of PCR
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Southern
Blotting
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Northern Blotting
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Genetic markers
are sequences of DNA which have
been traced to specific locations on
the chromosomes and associated
with particular traits
it can be described as a variation
that can be observed
may be a short DNA sequence,
such as a sequence surrounding a
single-base pair change (single
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5 conditions of a suitable
DNA markers:
must be polymorphic
co-dominant inheritance
randomly and frequently
distributed throughout the
genome
easy and cheap to detect
reproducible
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Molecular markers
applications includes :
germplasm characterization
genetic diagnostics
Characterization of transformants
Study of genome
organization and phylogenetic
analysis
paternity testing and the
investigation of crimes
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RFLP
Restriction
Fragment length
polymorphsim involves
fragmenting a sample of DNA by
a restriction enzyme, which can
recognize and cut DNA wherever
a specific short sequence occurs
occurs when the length of a
detected fragment varies
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RFLP
advantages:
variant are co dominant
measure variation at the
level of DNA sequence, not
protein sequence
disadvantage:
AFLP
Amplified fragment length
polymorphsim in this analysis we
can amplify restricted fragments
and reduces the complexity of
material to be analyzed (approx
1000 folds)
it can be used for comparison by
which closely related species only
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AFLP
Advantages includes:
fast
relatively inexpensive
highly variable
Disadvantage:
markers are dominant
presence of a band could mean
the individual is either
homozygous or heterozygous for
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RAPD
Random amplification of polymorphic
DNA is a type of PCR reaction, but the
segments of DNA that are amplified are
random
Advantages:
Fast - relatively inexpensive
- highly
variable
Disadvantage:
Markers are dominant
presence of a band could mean the
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Micro satellite
polymorphism, SSR or
Simple sequence repeat
Disadvantage:
Relatively expensive and time consuming
to develop
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SNP
A
single-nucleotide
polymorphism (SNP) is a
DNA
sequence
variation
occuring when a single
nucleotide A, T, C or G- in
the genome (or other shared
sequence differs between
members of a species or
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STR
Principles of DNA
Isolation and Purification
DNA cam be isolated from any
nucleated cell
DNA is a giant anion in solution
Sources of DNA
- blood - Cultured cells (plant or
animal)
Bacteria - biopsies
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DNA isolation
is a routine procedure to
collect DNA for
subsequent molecular
analysis.
3 basic steps in DNA
extraction:
1. Cell disruption
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2.
3.
Basic Rules
blood first lyse (explode) the red
blood cells with a gentle detergent
such as Triton -X-100
Wash cell hemoglobin(and other
pigments) inhibits restriction
enzyme and TAQ polymerase
wear gloves to protect your
samples from you
autoclave all solutions and store in
fridge (except SDS and organic
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