Methods to study gene expression

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What is the purpose of using guanidinium thiocyanate (GTC) in RNA isolation?

To visualise RNA molecules on a nitrocellulose membrane

To remove rRNA from eukaryotic mRNA

To amplify mRNA for PCR

To inhibit the activity of RNases (correct)

What is the primary function of the Poly A+ selection step in eukaryotic mRNA isolation?

To amplify the target RNA molecule

To synthesise cDNA from mRNA

To remove genomic DNA from the RNA sample

To separate mRNA from rRNA (correct)

What is the purpose of the northern blotting technique?

To synthesize cDNA from mRNA

To amplify target RNA molecules

To isolate mRNA from eukaryotic cells

To visualise and quantify RNA molecules (correct)

Which of the following is a characteristic of reverse transcription-PCR?

Involves the synthesis of cDNA from mRNA Signup and view all the answers

What is the purpose of the exponential phase in PCR?

To amplify the target DNA sequence exponentially Signup and view all the answers

What is the function of SYBR Green in quantitative real-time PCR?

To preferentially bind to double-stranded DNA Signup and view all the answers

What is the purpose of using a dual-labelled probe in TaqMan PCR?

To release a fluorescent dye upon probe cleavage Signup and view all the answers

What is the purpose of making a cDNA library?

To clone and express eukaryotic genes in bacteria Signup and view all the answers

What is the purpose of ligating cDNA to a restriction enzyme-digested vector?

To create a recombinant vector for bacterial transformation Signup and view all the answers

What is the purpose of selecting for bacteria with a cDNA insert?

To identify bacteria that have taken up the recombinant vector Signup and view all the answers

What is the primary purpose of adding guanidinium thiocyanate (GTC) to the RNA isolation process?

To inhibit RNAses and prevent RNA degradation Signup and view all the answers

What is the primary benefit of using Northern analysis (blotting) over other RNA detection methods?

It allows for the visualization of RNA molecules and their size determination Signup and view all the answers

What is the primary function of the random hexamers in reverse transcription-PCR?

To act as a primer for the reverse transcription reaction Signup and view all the answers

What is the primary advantage of using quantitative real-time PCR over traditional PCR?

It allows for the real-time measurement of PCR product accumulation Signup and view all the answers

What is the primary purpose of creating a cDNA library?

To isolate and clone specific genes of interest Signup and view all the answers

What is the primary benefit of using oligo dT primers in reverse transcription-PCR?

They allow for the amplification of RNA molecules with poly A+ tails Signup and view all the answers

What is the primary purpose of the linear phase in PCR?

It is the stage where the PCR product accumulates at a constant rate Signup and view all the answers

What is the primary benefit of using gene-specific primers in reverse transcription-PCR?

They enhance the specificity of the PCR reaction Signup and view all the answers

What is the primary purpose of transferring the RNA molecules to a nitrocellulose membrane in Northern analysis?

To immobilize the RNA molecules for hybridization with a probe Signup and view all the answers

What is the primary benefit of using reverse transcription-PCR over traditional PCR?

It allows for the analysis of RNA expression levels Signup and view all the answers

What is the primary reason why RNA isolation difficulties often arise?

Presence of RNases in the sample Signup and view all the answers

What is the primary advantage of using Poly A+ selection in eukaryotic mRNA isolation?

It helps to remove rRNA from the sample Signup and view all the answers

What is the primary purpose of transferring the RNA molecules to a nitrocellulose membrane in Northern analysis?

To immobilize the RNA molecules Signup and view all the answers

What is the primary function of the quencher in a TaqMan probe?

To prevent fluorescence when the probe is intact Signup and view all the answers

What is the primary advantage of using quantitative real-time PCR over traditional PCR?

It allows for the measurement of PCR products in real-time Signup and view all the answers

What is the primary purpose of making a cDNA library?

To study gene expression Signup and view all the answers

What is the primary function of the random hexamers in reverse transcription-PCR?

To prime the synthesis of cDNA Signup and view all the answers

What is the primary advantage of using reverse transcription-PCR over traditional PCR?

It allows for the analysis of RNA molecules Signup and view all the answers

What is the primary purpose of ligating cDNA to a restriction enzyme-digested vector?

To clone the cDNA Signup and view all the answers

What is the primary function of the exponential phase in PCR?

To amplify the target DNA sequence Signup and view all the answers

Methods to study gene expression

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Questions and Answers

What is the purpose of using guanidinium thiocyanate (GTC) in RNA isolation?

What is the primary function of the Poly A+ selection step in eukaryotic mRNA isolation?

What is the purpose of the northern blotting technique?

Which of the following is a characteristic of reverse transcription-PCR?

What is the purpose of the exponential phase in PCR?

What is the function of SYBR Green in quantitative real-time PCR?

What is the purpose of using a dual-labelled probe in TaqMan PCR?

What is the purpose of making a cDNA library?

What is the purpose of ligating cDNA to a restriction enzyme-digested vector?

What is the purpose of selecting for bacteria with a cDNA insert?

What is the primary purpose of adding guanidinium thiocyanate (GTC) to the RNA isolation process?

What is the primary benefit of using Northern analysis (blotting) over other RNA detection methods?

What is the primary function of the random hexamers in reverse transcription-PCR?

What is the primary advantage of using quantitative real-time PCR over traditional PCR?

What is the primary purpose of creating a cDNA library?

What is the primary benefit of using oligo dT primers in reverse transcription-PCR?

What is the primary purpose of the linear phase in PCR?

What is the primary benefit of using gene-specific primers in reverse transcription-PCR?

What is the primary purpose of transferring the RNA molecules to a nitrocellulose membrane in Northern analysis?

What is the primary benefit of using reverse transcription-PCR over traditional PCR?

What is the primary reason why RNA isolation difficulties often arise?

What is the primary advantage of using Poly A+ selection in eukaryotic mRNA isolation?

What is the primary purpose of transferring the RNA molecules to a nitrocellulose membrane in Northern analysis?

What is the primary function of the quencher in a TaqMan probe?

What is the primary advantage of using quantitative real-time PCR over traditional PCR?

What is the primary purpose of making a cDNA library?

What is the primary function of the random hexamers in reverse transcription-PCR?

What is the primary advantage of using reverse transcription-PCR over traditional PCR?

What is the primary purpose of ligating cDNA to a restriction enzyme-digested vector?

What is the primary function of the exponential phase in PCR?

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