Biotechnology and Molecular Biology Flashcards
Biotechnology
Biotechnology is the exploration of biological processes for industrial and other purposes, especially the genetic manipulation of microogranisms for the production of antibiotics and hormones
Molecular biology
Molecular biology is the study of biological organisms at the molecular level
- Use of biological systems to make desired end products
- Can use whole organisms or components of an organism (enzymes)
- Does not require genetically engineered organisms
Biotechnology
- Intentional modification of genomes for practical purposes
- Deliberate alteration of genotype to change phenotype
- Created using biotechnology and molecular biology tools
Genetic Engineering
- The study of organisms at the molecular level
- Includes genetics, biochemistry, microbiology, biotechnology, and genetic engineering\
- Relies on the use of DNA and RNA and their associated enzymes
Molecular Biology
What are the tools of molecular biology?
Tools of Molecular Biology
- PCR
- Gel Electrophoresis
- DNA microarrays
- Recombinant DNA
- Blots
- CRISPR System/ Cas 9
PCR stands for?
Polymerase Chain Reaction
In vitro DNA replication
- Can “amplify” small amounts of DNA into lots of DNA
- Can detect specific genes
- Can be used to clone or copy specific sequences
- Used in medicine, forensics, empirical research
PCR
- Uses automated equipment (thermocycler) to run through several replication cycles
- Each replication cycle doubles the amount of DNA produced
- Technology possible due to Taq polymerase, a heat stable DNA Polymerase
PCR
- Consists of three steps
- Length of and the number of cyles varies
- Reaction mixture contains target DNA, A,T,C,G, primers, enzymes, and buffers
PCR Cycle
What are the three steps of the PCR Cycle
PCR Cycle
- Denaturation
- Priming (annealing)
- Extension
- Heat is used to separate double strands of the DNA molecule so existing strands can be used as templates
- Hydrogen bonds break at ~95 degrees celsius
- Mimics activity of Helicase
Denaturation
- First step in PCR cycle
- DNA primers from the hydrogen bonds to the target sequence
- The primers are short (~25 bp) of synthetic pieces of DNA that are complementary to the target sequence of DNA
- One primer is required for each template strand
- Specific primers have complementary bases (correlates to specific genes)
- Priming step usually around 55 to 65 degrees celsius
- Mimics activity of primase
Priming (Annealing)
- Second step of PCR Cycle
- Taq polymerase (DNA polyermase) replicates template DNA
- Named after the source organism Thermus aquaticus
- Temperature usually at 72 degrees Celsius
- Can copy 1000 bp in 10 seconds (100 bp/sec)
- Times vary based on length of piece of DNA being copied
Extension
- Third and final step of PCR Cycle
- Technique used to analyze pieces of nucleic acids (DNA or RNA) that uses a gel matrix and electricity
- Gel is made from agarose, the purified sugar from agar
- DNA and RNA are negatively charged (phosphate groups), and move towards a positive charge
- When set up, nucleic acids travel through gel, and are separated by size
Gel Electrophoresis
PCR
Denaturation
- First step of PCR Cycle
- ~95 degrees celsius to break hydrogen bonds
- Mimics
- Helixase
Priming (Annealing)
- Second step of PCR Cycle
- 55 to 65 degrees Celsius
- Mimics primase
Extension
- Third and final step of PRC Cycle
- Temperature 72 degrees Celsius
- Mimics DNA polymerase
- Uses Taq polymerase
Gel Electrophoresis
Gel Electrophoresis
- Smaller DNA/RNA fragments migrate faster and therefore farther from the starting point
- The size of the DNA/RNA fragments can be determined by comparing strands
- Is a routine procedure used to visualize PCR products, samples of mRNA from cells, quality of isolated DNA
Gel Electrophoresis
- Process brings with gel electrophoresis and tranfers DNA/RNA fragments to a fancy piece of paper
Blots