NGS and mRNA isolation

What does the transcriptome represent in a cell or tissue?

• The entire set of DNA sequences

• The genes actively being expressed at a given time

• The complete set of proteins

• The metabolic state of the organism

What does the metabolome include?

• Only mRNA and non-coding RNAs

• Only enzymes and structural proteins

• Metabolic intermediates, hormones, signaling molecules, and other small molecules

• Only DNA sequences and regulatory elements

What is the primary role of TRIzol reagent in RNA isolation?

• To wash the RNA pellet

• To precipitate RNA from the aqueous phase

• To dissolve the purified RNA

• To lyse cells and inactivate RNases

Which of the following correctly describes the role of chloroform in RNA isolation?

• It facilitates phase separation, isolating RNA in the aqueous phase

• It dissolves the RNA pellet for storage

• t precipitates RNA from the aqueous phase

• It inactivates RNases in the sample

Why is isopropanol used during the RNA isolation process?

• To wash the RNA pellet

• To dissolve the RNA for storage

• To precipitate RNA by reducing its solubility

• To facilitate phase separation

During the phase separation step in RNA isolation, under acidic pH, where does the RNA predominantly remain?

• In the organic phase

• In the interphase

• In the isopropanol layer

• In the aqueous phase

What is the purpose of adding adapters during library preparation in Illumina sequencing?

• To label DNA with fluorescent tags

• To sequence longer DNA fragments

• To provide sequences needed for attaching DNA to the flow cell

• To remove non-bound DNA fragments

Which of the following best describes the function of bridge amplification in Illumina sequencing?

• To denature DNA and form single strands

• To create clusters of identical DNA sequences for efficient sequencing

• To cleave DNA into shorter fragments

• To detect fluorescent signals from incorporated nucleotides

Which of the following would be filtered out during the quality control step in Illumina sequencing?

• Reads with fluorescent signals

• Reads that match the reference genome perfectly

• Reads from a single DNA fragment

• Reads from clusters that overlap or have low intensity

What is meant by "coverage" in the context of NGS?

• The percentage of fluorescent signals detected

• The number of reads for each nucleotide

• The extent to which the target DNA is fully sequenced

• The number of index sequences used in sequencing