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EB 3410 Chapter 1
Biotechnology: Science and Technology AND In-Fusion Bio Brick Assembly Paper
| Question | Answer |
|---|---|
| Biotechnology: The technological application that uses biological systems, living organisms, or derivatives thereof, to __________, as well as to _____________ in industrial processes for specific uses. | Solve problems; make or modify useful products |
| What are 4 categories in industry for Biotechnology? | (1)Life Science (2)Agriculture (3)Food (4)Medicine |
| Why is biotechnology called a revolution now? | subcategories are now defined |
| Biotechnology is not a pure scientific discipline. What are the sub-fields of the biological sciences? | Genetics, Molecular Biology, Biochemistry, Cell Biology, and Microbiology |
| Biotechnology is not a pure scientific discipline. What are the sub-fields of the non-biological sciences? | Engineering, chemistry, physics, information technology |
| What are the parts of a biotechnology toolkit? | Biological; chemical; instrumentation; software |
| What are the 3 main subsets of biotechnology? | Agri-biotech., Healthcare biotech, and Industrial biotech. |
| What are Bio Bricks? | Sequences/Genes used to make a genetic circuit |
| What is the composition of Bio Bricks when purchased? | lyophilized DNA in a 384-wel plate |
| Where is the physical DNA stored when a Bio Brick is purchased? | DNA sequence on a plasmid |
| How are Bio Brick assembled (standard) | 2 plasmids, RE enzyme digest, leaves scar |
| How are Bio Bricks assembled (alternative) | PCR; purification; In-Fusion reaction |
| What is a scar sequence? | an 8 bp sequence (that replaces the RE binding sites); located between the two DNA sequences |
| What is special about the in-Fusion enzyme? | It has exonuclease activity, finds linear DNA with overhangs to connect the linear DNA strand into a circular plasmid |
| What are the advantages of the Clontech In-Fusion PCR cloning kit? | it is faster, more flexible, and has a higher success rate than the standard assembly |
| What are the disadvantages of the Clontech In-Fusion PCR Cloning kit? | It is more expensive, requires custom primers, and occassionally there are mutations in the plasmids |
| What is the drive behind In-Fusion BioBrick assembly? | improve the success rate//create new genes -- More Possibilities |
| What is Phusion Taq? | DNA Polymerase: very low error rate, more accurate, faster, obtain more product/amount of enzyme used, loading dye is included |
| How much smaller is the error rate of Phusion Taq to other Taq polymerase? | 50-fold smaller |
| Why is the PCR reaction digested with DpnI? | Recognizes and degrades methylated DNA- in this case, it will degrade the bacterial DNA (and leave the genes for the genetic circuit) |
| Why is standard assembly used more often than custom DNA synthesis? | custom DNA synthesis is still too expensive for most synthetic biology labs |
| What are the advantages of Clontech In-Fusion assembly OVER standard assembly | bio bricks might not be acessible yet (also, flexible, fast, and high success) |
Created by:
savelae
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