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Bio Exam II
DNA Tech
| Term | Definition |
|---|---|
| PCR: Polymerase Chain Reaction | - generates a lot of copies of DNA in short amount of time - Important when studying genes |
| PCR Components | - Template DNA - primers - dNTPs ( dideoxynucleosides) A, T, C, G - Taq. polymerase |
| Process of PCR- done in thermal cycle to modulate temp. of mixture | 1) denature: heat to 90 degrees C , breaks h-bonds in double helix 2) annealing: 40-65 degrees C, primers bind w/ complementary sequence in template stand 3) Extension: Taq. polymerase replicates DNA from 3'-5' end of primers. Strand become template |
| Gel Electrophoresis | |
| Agarose Gel | - polysaccharide gel - porous - passes electrical current through causing DNA to move from - to + end - Uses: confirms successful PCR, DNA sequencing, DNA profiling, Disease Diagnosis |
| Deoxyribonucleotide chain termination sequencing | - 1st automated sequencing method - Sanger: won Nobel prize in 1980 -can sequence DNA fragments to about 1000 bp. - same process as PCR but w/ one major diff |
| Ingredients for Deoxyribonucleotide chain termination squencing | - target dna - primer - DNA pol. -4 deoxyribonucleotides -4 dideoxyribonucleotides |
| Dideoxyribonucleotides | - missing the 3' hydroxyl group making in responsible for chain termination because the phosphate cant bind |
| Process of Deoxyribonucleotide chain termination squencing | - target DNA denatured - complementary strand synthesized - deoxyribonucleotide randomly incorporated - elongation stops (termination) at dideoxyribonucleotide |
| Polyacrylamide Gel | - made from polymer - DNA fragments put in get and they migrate based on size (shorter strands move faster) - Detector reads bases as they migrate |
| Uses of sequencing | - evolutionary relationships - determine loci & function - disease diagnosis |
| Applications | - species diversity: know organisms in community, otherwise very hard to measure small & rare organisms - DNA bar coding: sequence of small region of genome to identify species |
| Gene Therapy | - FDA approved gene therapy to cure type of genetic blindness; uses a retrovirus to insert correct genes into cell of retina |
| CRISPR | - silence or insert genes |
Created by:
tennadeleta.
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