The document discusses using plants as bioreactors to produce and extract DNA, RNA, and proteins. It describes the principles and applications of various molecular biology techniques including DNA extraction, polymerase chain reaction (PCR), plant transformation, tissue culture, restriction fragment length polymorphism (RFLP), Southern blotting, RNA extraction, gel electrophoresis, Northern blotting, protein extraction via polyacrylamide gel electrophoresis (PAGE), and Western blotting. These techniques can be used to analyze genetic diseases, produce copies of genes of interest, create disease resistant plants, and express proteins for applications such as synthetic hormones, antibodies, and artificial organs.

1. PLANTS AS
BIOREACTORS
DNA extraction
• Hina Amir
• Shakiba Anwar
• Sadaf Moneeba

2. 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

3. PRINCIPLE
DNA Isolation
• Removal and separation of
DNA.
1. Cell Lysis
2. Precipitation
3. Wash
4. Resuspension

4. • human blood
• DNA
TOPIC
APPLICATIONS
DNA sequencing
Check genetic diseases
PCR template
Analyze forensic evidence

5. PRINCIPLE
Polymerase Chain Reaction
Exponential amplification sequence of interest
linked to replication in vivo

7. APPLICATIONS
TOPIC
• Gene
•
COPIES
OF
GENE
Direct cloning from genomic DNA or cDNA
Analyze forensic samples
 Increase amount of DNA

8. PRINCIPLE
Plant Transformation
integration (foreign DNA ) cell regeneration
Expressing gene of interest

9. APPLICATIONS
TOPIC
• Gene of interest
• vector
Disease free plants
 Improve quality of plants (salt
resistance, drought resistant plants)
Hormone expression
pest resistance

10. PRINCIPLE
Tissue Culture
cell or tissue or organ fully differentiated organism

11. APPLICATIONS
TOPIC
• Transformed
Tobacco cells
Synthetic hormones
 Antibodies
Artificial pancreas —islet cells
Artificial bladders
Cartilage.
Artificial skin

12. PRINCIPLE
DNA Isolation

13. TOPIC
• DNA from cell
cultured plant

14. PRINCIPLE
Restriction fragment length
polymorphism

15. APPLICATIONS
TOPIC
• Restriction
fragments of
required
length
Paternity cases
Criminal cases
Disease status of an
individual

16. PRINCIPLE
Southern Blotting

17. APPLICATIONS
TOPIC
Amount of gene
or DNA
 DNA profiling
 Proving someone guilty
or innocent
Paternity Testing

18. PRINCIPLE
RNA Isolation
1. Cells or tissue disruption
2. Inactivate RNases
3. Denature nucleic acid-protein
complexes
4. RNA isolated
TOPIC
• RNA from plant

20. PRINCIPLE
Gel electrophoresis TOPIC
• Proteins from plant
• Migration of charged
molecules in an electric
field through a solution
or solid support
• Polyacrylamide –
Proteins
• Good preparative and
analytical method

22. PRINCIPLE and APPLICATIONS TOPIC
• Northern blotting
Sample preparing.
Transfer.
Adding probe.
Incubation.
Blot visualization
Gene's expression
pattern

24. PRINCIPLE and APPLICATIONS
TOPIC
• Proteins Adding SDS.
 heat.
 Transfer of denature
protein.
 Loading sample.

25. SDS-PAGE

26. PRINCIPLE and APPLICATIONS
TOPIC
• ProteinsSample preparation.
Transfer.
Incubation.
Adding antibody.
Confirmatory test in HIV.

28. 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