Lab Final Flashcards
What primers were used in the GMO lab?
tubulin and 35S
What is a 35S promoter? How many PCR primers are used to amplify this region?
35S promoter drives the expression of glyphosate-resistance, and one primer the 35S primer
What is the purpose of identifying tubulin?
Tubulin is present in all plants. Identifying tubulin proves you were looking at a plant product.
Why did we test Round-Up Ready soy and Wild Type soy products?
Round-Up Ready is genetically modified and wild type soy plants are not genetically modified. Those were the positive and negative controls.
What was the length of the primers used? Why is this a good length?
The primers were about 25 letters long. This is a good length because it is less likely for the primer to bind to non-target DNA
What stains could have been used on the gel?
carolinaBLU, CoomassieBlu, Ethydium Bromyde,
Why is the Drosophila Melanogaster a good model organism?
easy to keep in lab, short generation time, easy to see inherited traits, 8 chromosomes
Why are we looking at the chromosomes found in the salavary gland?
The chromosomes found in the salavary gland could be seen with a compound microscope after fixation
What are polytene chromosomes?
over-sized chromosomes that have developed from normal chromosomes
Why do the chromosomes have a banding pattern? What are the light and dark regions?
The banding pattern is visible landmarks to identify the location of a gene. The dark bands are whrer DNA is tightly packed (heterochromatin) and light bands are where it is less densely packed (euchromatin)
Why is there a “puff” sometimes in the chromosomes?
The “puff” is where the DNA has unwound its self to make it accessible to translation machinery
What in a drosophilia cell?
8 chromosomes, 4 pairs
What is the purpose of CaCl?
CaCl is to
What role does IPTG play in expressing GFP?
When the lac repressor binds to the promoter T7 polymerase is blocked & GFP cannot be expressed. When IPTG is added the lac repressor is inactivated and T7 is expressed.
How is GFP used as a marker gene?
GFP is tagged to a specific gene and when the gene is expressed so is GFP
What is the purpose of the heat shock?
The heat shock opens the pores in the bacterial cell wall so the vector can enter
What would happen if cells weren’t placed on ice immediately after the heat shock?
It would be possible the vector could escape the cell because the cell wall would be open too long
Why were plates incubated at 37C?
Plates were incubated to allow cell wall recovery and growth of the bacteria
What was the purpose of all four bacterial plates?
-DNA on LB plate: +control, show the bacteria will grow
-DNA on AMP plate: -control, show it doesnt have AMPR
+DNA on +AMP: grow bacteria on AMP
+DNA on +AMP/+IPTG: grow bacteria, glowing colonies
What can influence transformation efficiency?
recovery time, cross-contamination, shock wasn’t preformed quick enough
Name 2 arguments for the use of genetically modified crops
- crop yield
2. insect resistance
Name 2 arguments against the use of genetically modified crops
- unnatural
2. may cause super wees or kill off non target insects
What compounds are needed for PCR
h20, taq, primers, buffer, mgcl2, dNTPs
