Cloning and Genetic Tools Flashcards
(31 cards)
What is recombinant DNA?
DNA formed by combining DNA from two different organisms.
What is genetic engineering?
The alteration of genetic material to obtain desired traits, often involving recombinant DNA.
List applications of recombinant DNA technology.
Agriculture, forensic science, medical research, human genome project, vaccine production, gene function studies.
What careers are available in molecular biotechnology?
Lab-based: Scientist, Lab Manager, Technician; Non-lab: Educator, Project Leader, Biostatistician, Sales.
What does cloning refer to in biotechnology?
Creating exact genetic copies of DNA fragments, cells, or organisms.
Do clones occur naturally?
Yes; via asexual reproduction or identical twins.
Give examples of animal cloning.
Dolly the sheep (1996), Zhong Zhong and Hua Hua monkeys (2018).
What is therapeutic cloning?
Cloning cells to grow transplantable organs.
What is a vector in gene cloning?
A DNA segment like a plasmid used to carry foreign DNA into a host.
What are plasmids?
Small, circular DNA molecules used in genetic engineering.
Outline steps in genetic engineering using plasmids.
Identify GOI, amplify with PCR, integrate into plasmid, transform into host, validate.
How does PCR work?
It amplifies DNA using primers and polymerase through denaturation, annealing, and extension.
What are essential components of a PCR?
Template DNA, primers, DNA polymerase, buffer, dNTPs.
What is the function of a thermocycler?
It controls temperature cycles during PCR.
How should primers be designed?
18–24 bases long, 40–60% GC, Tm 50–60°C, avoid secondary structures.
What are restriction endonucleases?
Enzymes that cut DNA at specific sequences, creating sticky ends.
What is DNA ligase used for?
It joins DNA fragments with complementary sticky ends.
What are multiple cloning sites (MCS)?
Plasmid regions with several restriction sites for inserting genes.
How are restriction sites added to PCR products?
Integrated via primers’ 5′ ends.
What features do expression plasmids have?
Self-replication, selectable marker, regulatory sequences, inducible expression.
How is heat-shock transformation used in E. coli?
To introduce plasmid DNA into bacterial cells.
What is gel electrophoresis used for?
To check DNA size and validate cloned plasmids.
What product size is expected in PCR if an 800bp insert is added to a 150bp MCS?
Approx. 950bp.
