MODERN RESEARCH TECHNIQUES (BIT-319) Credit Hours 3(2-1) Educational Objectives: This particular...
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MODERN RESEARCH TECHNIQUES (BIT-319) Credit Hours 3(2-1) Educational Objectives: This particular course is designed to provide students understanding of
MODERN RESEARCH TECHNIQUES (BIT-319) Credit Hours 3(2-1)
Educational Objectives: This particular course is designed to
provide students understanding of basic concepts of research and
its methodologies. The students will learn about modern techniques
used in the field of applied biosciences. This will develop
research interest and will help them identify local problems and
finding the appropriate solutions using biotechnological
procedures. PRE- REQUISITE(s): Thorough knowledge of Microbiology,
Vaccinology and Virology is required. MODERN RESEARCH TECHNIQUES
(BIT-319) Credit Hours 3(2-1) Educational Objectives: This
particular course is designed to provide students understanding of
basic concepts of research and its methodologies. The students will
learn about modern techniques used in the field of applied
biosciences. This will develop research interest and will help them
identify local problems and finding the appropriate solutions using
biotechnological procedures. PRE- REQUISITE(s): Thorough knowledge
of Microbiology, Vaccinology and Virology is required.
Slide 2
Course Contents 1.DNA and RNA isolation 2.Quantification of DNA
and RNA 3.Primer designing 4.PCR 5.Electrophoresis 6.Sequencing
7.Karyotyping 8.Restriction Mapping Course Contents 1.DNA and RNA
isolation 2.Quantification of DNA and RNA 3.Primer designing 4.PCR
5.Electrophoresis 6.Sequencing 7.Karyotyping 8.Restriction Mapping
9.Flow cytometry 10.Hybridization a.Western blotting b.Southern
blotting c.Northern blotting d.FISH 9.Flow cytometry
10.Hybridization a.Western blotting b.Southern blotting c.Northern
blotting d.FISH
Slide 3
11.Transfection 12.Transduction 13.Transformation 14.Cloning
15.Microarrays 16.Chromatography 17.Immunochemistry 18.ELISA
19.Bioinformatics and techniques 20.Ethical issues 11.Transfection
12.Transduction 13.Transformation 14.Cloning 15.Microarrays
16.Chromatography 17.Immunochemistry 18.ELISA 19.Bioinformatics and
techniques 20.Ethical issues 21.Tissue culturing 22.Slide
Preparation and Cell Stains 23.Agar plate preparation and streaking
for the purpose of individual colony isolation 24.Bacterial Growth
on selective agar 25.Quantification: Colony Forming Units (CFU)
26.Dilution Plating 27.Identification and characteristics of
colonies 21.Tissue culturing 22.Slide Preparation and Cell Stains
23.Agar plate preparation and streaking for the purpose of
individual colony isolation 24.Bacterial Growth on selective agar
25.Quantification: Colony Forming Units (CFU) 26.Dilution Plating
27.Identification and characteristics of colonies
Slide 4
Lab Work: DNA extraction Quantification PCR Electrophoresis
ELISA Agar plate preparation Streaking Transfection Cloning Lab
Work: DNA extraction Quantification PCR Electrophoresis ELISA Agar
plate preparation Streaking Transfection Cloning
Slide 5
Recommended Books: Immunoassay and other bioanalytical
techniques by Eman.J.Immunoassay Nano biotechnology by C.M.
Niemeyer, C.A. Mirkin MLT (medical laboratory technology): methods
and interpretation by Rammiksood Genetic techniques for biological
research: a case study approach by Corinne V. Anthony
MichelsGenetic techniques for biological research: a case study
approach Corinne V. Anthony Michels Research techniques in
biochemistry and molecular biology by Robert E. Thach, Mary R.
NewburgerResearch techniques in biochemistry and molecular
biologyRobert E. ThachMary R. Newburger Methods in molecular
biology and protein chemistry: cloning and characterization of an
enterotoxin subunit by Brenda D. Spangler.Brenda D. Spangler
Recommended Books: Immunoassay and other bioanalytical techniques
by Eman.J.Immunoassay Nano biotechnology by C.M. Niemeyer, C.A.
Mirkin MLT (medical laboratory technology): methods and
interpretation by Rammiksood Genetic techniques for biological
research: a case study approach by Corinne V. Anthony
MichelsGenetic techniques for biological research: a case study
approach Corinne V. Anthony Michels Research techniques in
biochemistry and molecular biology by Robert E. Thach, Mary R.
NewburgerResearch techniques in biochemistry and molecular
biologyRobert E. ThachMary R. Newburger Methods in molecular
biology and protein chemistry: cloning and characterization of an
enterotoxin subunit by Brenda D. Spangler.Brenda D. Spangler
Slide 6
DNA ISOLATION Why to isolate where to isolate
Slide 7
1- Cell lysis 1.Sonication, 2.vortexing with pheno(cell walls n
capsids) 3.SDS for lipid 4.protease. DNA associated+cellular
proteins 2- Protein Precipitation a.salt ammonium or sodium
acetate. b.phenol-chloroform: vortexed n entrifuged c.proteins will
remain in the organic phase and can be drawn off carefully. MANUAL
How to isolate
Slide 8
DNA precipitation: mixing with cold ethanol or isopropanol and
then centrifuging. alcohol serves as a wash to remove the salt
previously added. DNA precipitation: mixing with cold ethanol or
isopropanol and then centrifuging. alcohol serves as a wash to
remove the salt previously added. Washing: the resultant DNA pellet
with cold alcohol again and centrifuge for retrieval of the pellet.
Washing: the resultant DNA pellet with cold alcohol again and
centrifuge for retrieval of the pellet. Drying: the pellet, the DNA
can be re-suspended in a buffer such as Tris or TE. Drying: the
pellet, the DNA can be re-suspended in a buffer such as Tris or TE.
Visualization: Presence of DNA can be confirmed by electrophoresing
on an agarose gel containing ethidium bromide, or another
fluorescent dye that reacts with the DNA, and checking under UV
light Visualization: Presence of DNA can be confirmed by
electrophoresing on an agarose gel containing ethidium bromide, or
another fluorescent dye that reacts with the DNA, and checking
under UV light
Slide 9
PH 7= DNA +RNA in aqueous PH 4= DNA precipitated PH 7= DNA +RNA
in aqueous PH 4= DNA precipitated
Slide 10
Slide 11
Chelating agent to (EDTA) sequester divalent cations such as,
Mg 2+ and Ca 2+, which prevents enzymes like DNase from degrading
the DNA.