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DNA extraction (human)
PCR (gene of interest)
Transformation
Tissue Culture
Southern blotting Northern blotting Western blotting
DNA extraction and RFLP
RNA extraction and gel electrophoresis
Protein extraction and PAGE
PRINCIPLE
DNA Isolation
• Removal and separation ofDNA.
1. Cell Lysis
2. Precipitation
3. Wash
4. Resuspension
• human blood
• DNA
TOPICAPPLICATIONS
DNA sequencingCheck genetic diseasesPCR templateAnalyze forensic evidence
PRINCIPLE
Polymerase Chain Reaction
Exponential amplification sequence of interestlinked to replication in vivo
APPLICATIONSTOPIC
• Gene
•
COPIESOF
GENE
Direct cloning from genomic DNA or cDNA
Analyze forensic samples
Increase amount of DNA
PRINCIPLE
Plant Transformation
integration (foreign DNA ) cell regeneration
Expressing gene of interest
APPLICATIONSTOPIC
• Gene of interest
• vector
Disease free plants Improve quality of plants (saltresistance, drought resistant plants)Hormone expressionpest resistance
APPLICATIONSTOPIC
• Transformed Tobacco cells
Synthetic hormones AntibodiesArtificial pancreas —islet cellsArtificial bladders Cartilage.Artificial skin
APPLICATIONSTOPIC
• Restriction fragments of required length
Paternity cases
Criminal cases
Disease status of an
individual
APPLICATIONSTOPIC
Amount of gene or DNA
DNA profiling
Proving someone guilty or innocent
Paternity Testing
PRINCIPLE
RNA Isolation
1. Cells or tissue disruption2. Inactivate RNases3. Denature nucleic acid-protein
complexes4. RNA isolated
TOPIC
• RNA from plant
PRINCIPLE
Gel electrophoresisTOPIC
• Proteins from plant• Migration of charged
molecules in an electric field through a solution or solid support
• Polyacrylamide –Proteins
• Good preparative and analytical method
PRINCIPLE and APPLICATIONS TOPIC
• Northern blottingSample preparing.Transfer.Adding probe. Incubation.Blot visualization
Gene's expression pattern
PRINCIPLE and APPLICATIONS TOPIC
• Proteins Adding SDS.
heat.
Transfer of denature protein.
Loading sample.
PRINCIPLE and APPLICATIONS TOPIC
• Proteins Sample preparation.
Transfer.
Incubation.
Adding antibody.
Confirmatory test in HIV.
REFERENCES
• MOLECULAR BIOLOGY LAB May 2010 Thoraia Shinawi
• Rodríguez-Lázaro, D. and Hernández, M. (2013). Introduction to the Real-time. Real-Time PCR in Food Science: Current Technology and Applications.Caister Academic Press, UK. ISBN: 978-1-908230-15-7.
• http://bioneer.com.au/Products/DNA-RNA-Preparation-Kits
• http://science.howstuffworks.com/dna-profiling1.htm
• http://www.bio.davidson.edu/genomics/method/RFLP.html
• http://www.highveld.com/pcr/principle-of-pcr.html
• http://www.jains.com/Tissue/images/tissue_main.jpg