1. Fill in the gaps that describes each step of PCR. (2 points each, 10 points total)
| Steps | Temperature | Mechanism (describe what is going on during PCR and include the name of enzymes if they're involved) |
|---|---|---|
| Denature | 1. ________ | 2. ________ |
| Anneal | 3. ____ - ____ | Oligonucleotide primers (forward and reverse) binds to their complement |
| 4. ________ | 72C | 5. ________ |
2. Design 6bp primers that amplify ONLY the underlined section. Label the 5' and 3' ends of the primers, indicate where these primers would anneal, and indicate in which direction elongation would occur (4 points)
5'-CTTACCTGTGCTGAGCATTAGACATGAACGGTTAGCTACTGAGCTAACA-3'
3'-GAATGGACACGACTCGTAATCTGTACTTGCCAATCGATGACTCGATTGT-5'
3. A widely used reporter gene in Arabidopsis thaliana plants (used in BIO253L) is the ______________ gene that encodes the enzyme GUS. This enzyme can cleave the chromogenic (color-generating) substrate ______________, resulting in the production of an insoluble __________ color in those plants. (6 points: abbreviated names for enzymes and substrates are OK)
