Biotechnology & synthetic bio

What is genetic engineering?

The manipulation of an organism’s genome.

What does biotechnology rely on?

Microorganisms and cells to make a product for human benefit.

What is synthetic biology?

Engineering biological systems by linking known components in various combinations.

How are genomes compared to software?

Genomes are programmable like software.

How are cells compared to hardware?

Cells function as factories, similar to hardware.

What are some applications of genetic engineering?

Medicine, agriculture, industry, and environmental protection.

What is the main difference between synthetic biology and traditional biotechnology?

Synthetic biology focuses on designing new biological systems while biotechnology modifies existing ones.

What is the central molecule manipulated in genetic engineering?

DNA.

What are plasmids?

Small circular DNA molecules that replicate independently and are used for gene manipulation.

What do restriction enzymes do?

They cut plasmid DNA at specific sequences to allow the insertion of foreign genes.

What is the role of DNA ligase in genetic engineering?

It seals the nicks in the sugar-phosphate backbone, facilitating the formation of recombinant DNA molecules.

What is PCR (Polymerase Chain Reaction)?

A pivotal technique that amplifies specific DNA sequences to generate numerous copies.

What are the applications of PCR?

Diagnosing genetic diseases, detecting mutations, detecting viruses, and isolating DNA segments.

What is required for PCR?

Target sequence, PCR DNA polymerase, and primers.

What temperature is typically used for denaturation in PCR?

94 degrees Celsius.

What is the extension temperature for Taq DNA polymerase in PCR?

72 degrees Celsius.

What does Reverse Transcription PCR (RT-PCR) do?

It converts mRNA into complementary DNA (cDNA) using reverse transcriptase.

What is agarose electrophoresis used for?

Separating DNA/RNA fragments by size.

What role do nucleic acid probes play?

They detect, characterize, and identify segments of DNA and RNA.

What is the purpose of Southern blotting?

To detect specific DNA sequences in a sample.

What does CRISPR-Cas system enable?

Precise editing of the genome, allowing for targeted modifications.

How does CRISPR utilize RNA?

It uses RNA to target specific DNA sequences for editing.

What defines a vector in genetic engineering?

A DNA molecule that carries DNA from one organism to another.

What is a selection marker in plasmids?

An antibiotic resistant gene that helps identify plasmid-containing colonies.

What are reporter genes used for?

To visualize DNA insertion in host cells.

What are the key steps in the workflow of cloning?

PCR to isolate gene, insert gene into vector, introduce vector into cells, select and express genes.

What is the aim of heterologous expression?

To express a gene from one organism in a different host organism.

What happens during transformation in genetic engineering?

Bacteria take up free DNA from the environment.

What is the purpose of using electrocompetent cells?

To enhance the uptake of DNA by bacteria during transformation.

What does blue/white screening indicate in cloning?

It differentiates between colonies with and without foreign DNA.

What does the enzyme reverse transcriptase do?

It synthesizes cDNA from an RNA template.

What is hybridization in molecular biology?

The process where single-stranded nucleic acid binds to its complementary sequence.

What does FISH stand for in genetic research?

Fluorescence In Situ Hybridization.

What is a common challenge with CRISPR technology in human genome editing?

Even though it is specific, there is still a chance of off-target effects.

What can dideoxy nucleotides cause in DNA sequencing?

They terminate DNA synthesis during sequencing.

What information does DNA sequencing provide?

The order of nucleotides in a DNA fragment.

What is one main application of DNA sequencing?

To confirm successful genetic modification.

What safety measure is often taken when using CRISPR technology?

It is not used for human genome editing due to potential risks.

What is the significance of introns in gene expression?

Introns are present in eukaryotic genes but not in prokaryotic genes.

What role do agarose gels play in molecular biology?

They are used to separate nucleic acids during electrophoresis.

What do fluorophores in probes allow for in research?

They enable the visualization of specific nucleic acid sequences.

How does CRISPR provide a simpler method for genetic modifications?

It uses guide RNA to target specific DNA sequences, making editing more accurate.

Describe the process of hybridization in electrophoresis.

Nucleic acids can be denatured and then reannealed to form complementary sequences.

What can be achieved through recombinant DNA technology?

Producing medicines like insulin using bacteria.

What defines the difference between blue and white colonies in x-gal screening?

Blue colonies have intact lacZ and lack foreign gene; white colonies have interrupted lacZ and contain foreign gene.

How does PCR facilitate the diagnosis of diseases?

By amplifying specific DNA sequences to detect mutations or pathogens.

What is a common use case for plasmid vectors in research?

They are used to carry foreign DNA into target organisms.

What is the advantage of using Taq polymerase in PCR?

It is thermostable and allows for efficient DNA synthesis at high temperatures.

What is an essential outcome of using restriction enzymes?

They create sticky or blunt ends that facilitate DNA ligation.