pGLO Experiment SAC Flashcards
Independant Variable
The variable you change
Dependant Variable
The variable that changes basedd on the independant variable
Systematic Error
Error due to faulty equipment or calibration
Personal Error
Mistake/Miscalculation made by experimenter
Random Error
Errors caused by unpredictable and unaviodable variations
Control Variable
Variable that’s held constant through research
Polymerase Enzyme
An enzyme that builds a polymer
DNA Polymerase
Synthesies DNA in a 5’ to 3’ ratio using an existing strand as a template
Taq Polymerase
DNA polymerase taken from bacteria Thermus Aquaticus
Optimum functional temperature is 72 degress
Ligase
An enzyme that joins molecules , including DNA or RNA together, by catalysing the formation of Phosphodiester bonds
Endonuclease
Enzyme that cuts sugar-phosphate bonds between nucleotides
Palindrome
Sequence of double stranded DNA that reads the same on both strands in a 5’ to 3’ direction
eg:
5’ AGT|ACT 3’
3’ TCA|TGA 5’
Blunt Ends
Clean cut down the middle
eg:
AGC|ACT
TCG|TGA
Sticky Ends
Cut anywhere other than down the middle
GACGT|C
C|TGCAG
Polymerase
Adds nucleotides to DNA or RNA, which can lead to copying entire genes (amplifies sections)
Primer
Short, single strand of nucleic acids that acts as a starting point for polymerase enzyme to attach
Monomer
Molecule
What way does the polymerase enzyme read and synthesise complementary strands
in a 5’ to 3’ direction
Restriction Endonuclease
Cuts DNA/RNA at specific restriction sight
Why transform bacteria?
Genetically modifying bacteria to produce human proteins has revolutionised modern medicine and agriculture
Plasmid
Small, circular loop of DNA separate from the chromosome, typically found in bacteria
Recombinant Plasmid
Circular DNA vector that is ligated to incorporate a gene of interest
Bacterial Transformation
process by which bacteria take up foreign DNA from their environment. Used to introduce recombinant plasmids into bacteria
Genetic Modification
manipulation of an organism’s genetic material using biotechnology
Insulin
Hormone secreted by the pancreas to control blood glucose levels
Diabetes
Disease where the body cannot properly produce or respond to insulin
Vector
means of introducing foreign DNA into an organism. Plasmids are a popular vector in bacterial transformation
Antibiotic Resistance Gene
Gene which confers antibiotic resistance
Antibiotic
Destroys bacteria
Origin of Replication
Sequence found in prokaryotes that signals the start site of DNA replication
Reporter Gene
Gene with an easily identifiable phenotype that can be used to identify whether a plasmid has taken up a gene of interest
Heat Shock
Method of promoting recombinant plasmid uptake involving rapidly increasing and decreasing temperature to increase membrane permeability to enhance the likelyhood of bacterial transformation
Electroporation
Methods of promoting recombinant plasmid uptake that involves delivering an electric shock to bacterial membranes to increase membrane permeability and likelyhood of bacterial transformation
Fusion Protein
Protein made when separate genes have been joined and are transcribed and translated together
Restriction Enzymes
Cut DNA (6 base palindromic sequence)
Steps to Creating Recombinant Plasmids
1) Choose restriction enzyme
2) Choose a plasmid
3) Using restriction endonuclease
4) Making a recombinant plasmid
5) Transforming Bacteria
6) Identifying and Culturing Bacteria
Recombinant
Organism or DNA molecule containing DNA from more than one species
1) Choosing Restriction Enzyme
Endonuclease is chosen to cut upstream and downstram of gene, leaves stick ends
2) Choosing a plasmid
Plasmid is chosen that has 2 genes that each encode observable traits
Of these, one must contain the restriction site
3) Using Restriction Enzyme
Same restriction enzyme used to cut both source gene, and the plasmid
4) Making a recombinant plasmid
When source gene and plasmids are mixed, source gene may incorperate into plasmids, creating a recombinant
A plasmid containing foreign ‘passenger’ DNA is caleld ‘recombinant’
5) Transforming Bacteria
Bacteria made competent to take up plasmid
- only some bacteria will take up plasmid
- only some will take up RECOMBINANT plasmid
6) Identifying and Culturing Bacteria
Growing bacteria on agar plate can help determine whether the bacteria has taken up a plasmid
Bacteria that haven’t taken a plasmid want to be able to grow, though lack the resistance gene
Lawn
Covered in bacteria
Colonies
Small patches of bacteria
No growth
No bacteria - lacks resistance gene
Overhanging Nucleotide
unbonded nucleotides on the ends of DNA strand resulting from a staggered cut (sticky ends)
PCR
Polymerase Chain Reaction
Denaturation
Anneale
Elongation
Extraneous Variable
factor that is not accounted for, as is not kept constant through the experiment
Methodology
strategy followed in a scientific investigation
Method
the steps taken within an investigation
Accuracy
Results match the expectation for the experiment (may not be precise although)
Precision
Results are all close together, (may not be accurate although)
