Podcast
Questions and Answers
PCR is a laboratory technique for producing millions to billions of copies of RNA.
PCR is a laboratory technique for producing millions to billions of copies of RNA.
False (B)
PCR is also known as molecular photocopying.
PCR is also known as molecular photocopying.
True (A)
PCR is a slow and expensive technique.
PCR is a slow and expensive technique.
False (B)
The main activity of DNA polymerase is to cut DNA strands.
The main activity of DNA polymerase is to cut DNA strands.
Primers bind to the template RNA during PCR.
Primers bind to the template RNA during PCR.
PCR can be used to produce billions of copies of a specific segment of DNA for detailed study.
PCR can be used to produce billions of copies of a specific segment of DNA for detailed study.
Pfu polymerase is derived from Pyrococcus furiosus.
Pfu polymerase is derived from Pyrococcus furiosus.
Pfu polymerase has 5' to 3' exonuclease activity.
Pfu polymerase has 5' to 3' exonuclease activity.
Pfu polymerase is characterized by high processivity compared to Taq polymerase.
Pfu polymerase is characterized by high processivity compared to Taq polymerase.
1–2 units of Taq DNA polymerase are sufficient for a typical 50 µL PCR reaction.
1–2 units of Taq DNA polymerase are sufficient for a typical 50 µL PCR reaction.
Increasing the amount of DNA polymerase in a reaction with inhibitors may reduce PCR yields.
Increasing the amount of DNA polymerase in a reaction with inhibitors may reduce PCR yields.
Taq Polymerase has terminal transferase activity at room temperature.
Taq Polymerase has terminal transferase activity at room temperature.
Magnesium ions function as a cofactor for the activity of RNA polymerases.
Magnesium ions function as a cofactor for the activity of RNA polymerases.
Mg2+ facilitates the formation of complex between primers and DNA templates by destabilizing negative charges on their phosphate backbones.
Mg2+ facilitates the formation of complex between primers and DNA templates by destabilizing negative charges on their phosphate backbones.
Potassium ions from KCl promote primer annealing in PCR.
Potassium ions from KCl promote primer annealing in PCR.
Ammonium sulfate can replace KCl in the buffer to enhance specificity in PCR.
Ammonium sulfate can replace KCl in the buffer to enhance specificity in PCR.
Denaturation phase in PCR involves cooling the dsDNA template at 94-95 °C.
Denaturation phase in PCR involves cooling the dsDNA template at 94-95 °C.
Low Mg2+ concentrations in PCR result in enhanced stability of primer-template complexes.
Low Mg2+ concentrations in PCR result in enhanced stability of primer-template complexes.
Multiplex PCR involves amplifying multiple targets using a single primer pair.
Multiplex PCR involves amplifying multiple targets using a single primer pair.
Distinct amplicons in conventional PCR need to have similar base pair lengths for visualization.
Distinct amplicons in conventional PCR need to have similar base pair lengths for visualization.
Nested PCR increases the specificity of DNA amplification by reducing non-specific amplification of DNA.
Nested PCR increases the specificity of DNA amplification by reducing non-specific amplification of DNA.
A nested PCR assay typically has three sets of primers for a single locus.
A nested PCR assay typically has three sets of primers for a single locus.
The first PCR run in nested PCR uses the inner pair of primers.
The first PCR run in nested PCR uses the inner pair of primers.
Nested PCR results in the production of a longer PCR product compared to regular PCR.
Nested PCR results in the production of a longer PCR product compared to regular PCR.
During the extension step in PCR, DNA polymerase extends the primer sequences from the 5’ end of each primer.
During the extension step in PCR, DNA polymerase extends the primer sequences from the 5’ end of each primer.
A 2 min extension is typically sufficient to synthesize PCR fragments up to 3 kilobases (kb).
A 2 min extension is typically sufficient to synthesize PCR fragments up to 3 kilobases (kb).
During the first extension in PCR, the template will be length limiting, restricting the synthesis of templates exceeding the amplicon length.
During the first extension in PCR, the template will be length limiting, restricting the synthesis of templates exceeding the amplicon length.
A typical PCR reaction contains 20–30 cycles in a standard PCR run and can increase up to a maximum of 50 cycles.
A typical PCR reaction contains 20–30 cycles in a standard PCR run and can increase up to a maximum of 50 cycles.
Increasing the number of cycles in PCR with large amounts of starting material is advisable to ensure higher yield of PCR product.
Increasing the number of cycles in PCR with large amounts of starting material is advisable to ensure higher yield of PCR product.
At the end of a PCR reaction, amplification products are typically analyzed using mass spectrometry.
At the end of a PCR reaction, amplification products are typically analyzed using mass spectrometry.
Flashcards
PCR amplifies RNA
PCR amplifies RNA
PCR (Polymerase Chain Reaction) is a technique used to amplify specific DNA sequences, not RNA.
PCR is slow and expensive
PCR is slow and expensive
PCR is a powerful and versatile technique for amplifying DNA, making it a fundamental tool in molecular biology research.
DNA polymerase cuts DNA strands
DNA polymerase cuts DNA strands
DNA polymerase is an enzyme that adds nucleotides to a growing DNA strand, not cut them.
Primers bind to template RNA during PCR
Primers bind to template RNA during PCR
Signup and view all the flashcards
PCR produces copies of RNA
PCR produces copies of RNA
Signup and view all the flashcards
Pfu polymerase origin
Pfu polymerase origin
Signup and view all the flashcards
Pfu polymerase has 5' to 3' exonuclease activity
Pfu polymerase has 5' to 3' exonuclease activity
Signup and view all the flashcards
Pfu polymerase has high processivity
Pfu polymerase has high processivity
Signup and view all the flashcards
Taq polymerase units in a reaction
Taq polymerase units in a reaction
Signup and view all the flashcards
Increasing DNA polymerase reduces PCR yield
Increasing DNA polymerase reduces PCR yield
Signup and view all the flashcards
Taq polymerase has terminal transferase activity
Taq polymerase has terminal transferase activity
Signup and view all the flashcards
Mg2+ cofactor for RNA polymerase
Mg2+ cofactor for RNA polymerase
Signup and view all the flashcards
Mg2+ destabilizes phosphate backbones
Mg2+ destabilizes phosphate backbones
Signup and view all the flashcards
Potassium ions promote primer annealing
Potassium ions promote primer annealing
Signup and view all the flashcards
Ammonium sulfate replaces KCl in the buffer to enhance specificity in PCR
Ammonium sulfate replaces KCl in the buffer to enhance specificity in PCR
Signup and view all the flashcards
Denaturation phase involves cooling dsDNA
Denaturation phase involves cooling dsDNA
Signup and view all the flashcards
Low Mg2+ concentration results in increased stability of primer-template complexes
Low Mg2+ concentration results in increased stability of primer-template complexes
Signup and view all the flashcards
Multiplex PCR uses a single primer pair
Multiplex PCR uses a single primer pair
Signup and view all the flashcards
Distinct amplicons need similar base lengths to visualize
Distinct amplicons need similar base lengths to visualize
Signup and view all the flashcards
Nested PCR increases specificity by reducing non-specific amplification of DNA
Nested PCR increases specificity by reducing non-specific amplification of DNA
Signup and view all the flashcards
Nested PCR uses three primer sets for one locus
Nested PCR uses three primer sets for one locus
Signup and view all the flashcards
The first PCR run uses inner pair of primers
The first PCR run uses inner pair of primers
Signup and view all the flashcards
Nested PCR produces a longer PCR product
Nested PCR produces a longer PCR product
Signup and view all the flashcards
DNA polymerase extends primer sequences from the 5' end
DNA polymerase extends primer sequences from the 5' end
Signup and view all the flashcards
A 2 min extension is sufficient for 3kb fragments
A 2 min extension is sufficient for 3kb fragments
Signup and view all the flashcards
The template is not length limiting in the first extension
The template is not length limiting in the first extension
Signup and view all the flashcards
A typical PCR run contains 20-30 cycles and can go up to 50 cycles
A typical PCR run contains 20-30 cycles and can go up to 50 cycles
Signup and view all the flashcards
Increasing the number of cycles is advisable when starting material is high
Increasing the number of cycles is advisable when starting material is high
Signup and view all the flashcards
PCR products are analyzed using mass spectrometry
PCR products are analyzed using mass spectrometry
Signup and view all the flashcards