Biotechnology Principles & Processes By B.DEEPAN PhD
About myself Name: B.Deepan Qualification : M.Sc. PhD Experience: 10 years of experience Teaching NEET Biology ,NTSE Programme. Exams qualified: DBT exam, IELTS Vision & Mission: To make all my students (including you) as a doctor.
1.Taq Polymerase enzyme is obtained from 1.Thiobacillus ferroxidans 2.Bacillus subtilis 3.Pseudomonas subtilis 4.Thermus aquaticus
Hot temperature: Thermus aquaticu PCR -MACHINE
Extension of Nucleotides by DNA Polymerase enzyme dNTPs Primer 5 31 5 Template DNA New DNA strand DNA polymerase
3.DNA fragments generated by the restriction endonucleases in a chemical reaction can be separated by Polymerase chain reaction . Electrophoresis Restriction mapping Centrifugation
Electrophoresis- Instrument Power supply sample wells O electrode direction of movement electrode buffer solution Electrophoresis tank
Gel structure 1 2 3 bp 360 bp 500 400 300 200 232 bp 100
4.PCR and Restriction Fragment Length Polymorphism are the methods Study of enzyme:s Genetic transformation DNA sequencing . Genetic Fingerprinting for
Sealable Labeled plastic bag probe Filter paper Gel 1A 2The resulting 3 After electro- 4After 9 The fiter paperis To visualize 4 5 6 The filter paper is placed into a plastic bag containing a solution with the single-stranded labeled probe, and the filter paper is probe sticks to complementary fragments by hybrid- ization. (The bands are ive emissions not visible at this stage. fragments are separated by size by gel electrophoresis. restric- tion enzyme is added to a solution contain- ing the DNA of interest. phoresis, the DNA blotting, fragments in the gel are denatured paper is the positions of the bands where the probe hybridized, the the filter and then trans- removed ferred (blotted) onto filter paper. exposed to X-ray film, and either light or radioac- darken the film over each band.
DNA Ligase complementary sticky ends joins backbone (sugar-phosphate) GTTAA G TTAA AATTG Insert DNA Ligase Plasmid
.Agarose is made up of D-Galactose & 3,6- anhydro L-galactopyranose ring. . Agarose gel electrophoresis is a method to separate DNA /RNA according to their molecular size. Power supply O electrode of movement @ electrode buffer solution Electrophoresis tank * Smaller molecules move faster than the larger molecules
8. There is a restriction endonuclease called EcoRI. What does .co part in it stand for . Colon .Coenzyme Coelom Coli Strain Name 7 Escherichia Coli RY13 two letters of species name First letter of genus name
9. Bacillus thuringiensis forms protein crystals which contain insecticidal rotein .binds with epithelial cells of midgut of the insect is coded by several genes including the gene is activated by acid pH of the foregut of the insect does not kill the carrier bacterium which is itself pest ultimately killing it. CRY. pest. resistant to this toxin
10. DNA or RNA segment tagged with a radioactive molecule is called * Vector Probe . Clone . Plasmid
DNA Probe Denatured SSDNA DNA sample DNA probes gene of interest molecular beacon 1 Isolate DNA from 2 Denature DNA sample and combine with 3 DNA probes will bind body fluid sample. DNA probes. Probes are complementary to the gene of interest and labeled with a molecular beacon. to the gene of interest if it is present in the DNA sample.
MSC.Phd final year 10 years of Experience in teaching NEET Biology & NTSE. IELTS, DBT Exams Qualified