dna cloning

• What is the process called when bacteria take up foreign DNA?

  Transformation
• Why is transformation important in DNA cloning?

  It transfers plasmids to bacteria
• What are the two enzymes used in the ligation step of DNA cloning?

  Restriction enzymes and DNA ligase
• How are bacteria selected after transformation?

  They are placed on antibiotic plates
• What do bacteria with a plasmid become resistant to?

  Antibiotics
• What do colonies of bacteria represent after transformation?

  Clusters of identical, plasmid-containing bacteria
• Why do we need to check colonies after transformation?

  To ensure they contain the correct plasmid
• What can happen when inserting a gene into a plasmid?

  The plasmid may close without the gene
• What is the consequence of a gene being inserted backwards into a plasmid?

  No protein will be made
• What methods are commonly used to analyze plasmid DNA?

  Restriction digests, PCR, and DNA sequencing
• What happens if a colony contains a "good" plasmid?

  It can be used for plasmid or protein production
• How can bacteria be used as plasmid factories?

  They produce large amounts of plasmid DNA
• What is the role of control sequences in plasmids?

  They induce gene expression in bacteria
• What is the purpose of lysing bacteria in protein production?

  To release the produced protein
• What is affinity chromatography used for?

  To purify proteins from bacterial lysates
• What is the function of antibodies in affinity chromatography?

  They bind specifically to target molecules
• What happens to other molecules during affinity chromatography?

  They are washed away from the column
• What is the final step in affinity chromatography?

  The protein of interest is collected
• What is the purpose of the simulations from LabXchange mentioned in the material?

  To learn more about bacterial transformation
• What is the license type for the article mentioned in the material?

  CC BY-NC-SA 4.0
• What is the significance of the gene therapy clinical trial mentioned?

  It used plasmids to deliver a human gene
• What is the main goal of DNA cloning?

  To make many copies of a specific DNA piece
• What is a plasmid?

  A circular piece of DNA
• What is the role of heat shock in bacterial transformation?

  It makes the bacterial membrane more permeable
• What is the outcome for bacteria without a plasmid on antibiotic plates?

  They die
• What is the relationship between ligation and transformation?

  Ligation occurs before transformation
• How do plasmids contribute to gene expression in bacteria?

  They contain control sequences for expression
• What is the significance of checking plasmid DNA from colonies?

  To confirm the plasmid matches the intended design
• What is the first step in a typical cloning experiment?

  Insert DNA into a plasmid
• What happens to plasmids during the ligation process?

  They are joined with DNA fragments
• What is the purpose of using antibiotic plates in bacterial transformation?

  To select for bacteria that took up plasmids
• How does the heat shock method facilitate transformation?

  It creates pores in the bacterial membrane
• What is the role of DNA ligase in cloning?

  It joins DNA fragments together
• What is the typical outcome of a ligation reaction?

  Some good and some bad plasmids
• Why is it important to use the same restriction enzyme for cutting DNA?

  To ensure compatible ends for ligation
• What is the significance of the promoter in a plasmid?

  It drives gene expression in bacteria
• What happens to plasmids that do not contain the correct gene?

  They may close without the gene
• What is the purpose of using PCR in plasmid analysis?

  To amplify specific DNA sequences
• How does the presence of antibiotic resistance genes in plasmids aid selection?

  It allows for the survival of transformed bacteria
• What is the role of the immune system protein in affinity chromatography?

  It binds specifically to the target protein