Sanger Sequencing

What is the first step in designing primers for the Sanger method of DNA sequencing? (NIC)

• Selecting the DNA polymerase

• Identifying the sequence to be amplified

• Choosing the fluorescent dideoxynucleotides

• Running gel electrophoresis

Which enzyme is crucial for the Polymerase Chain Reaction (PCR) in the Sanger method?

• Ligase

• Restriction enzyme

• DNA polymerase

• RNA polymerase

What is the purpose of gel electrophoresis in the Sanger sequencing method?

• To amplify DNA fragments

• To cut DNA into smaller fragments

• To separate DNA fragments based on size

• To visualize the DNA directly

What role do fluorescent dideoxynucleotides play in the Sanger sequencing method? (NIC)

• They cut the DNA into smaller pieces.

• They amplify the DNA fragments.

• They terminate DNA synthesis and allow for detection.

• They bind to specific DNA sequences for identification.

Automated base calling in the Sanger sequencing method refers to: (NIC)

• Manually reading the sequence of bases from a gel

• Using a computer algorithm to determine the sequence of bases from fluorescent signals

• Calling out the bases as they are incorporated into the DNA strand

• Using a special phone app to record DNA sequences

Which component is not required for the Polymerase Chain Reaction (PCR) in the Sanger method?

• DNA template

• Primers

• RNA polymerase

• dNTPs

What is the main reason for using gel electrophoresis after the sequencing reaction in the Sanger method?

• To purify the DNA fragments

• To replicate the DNA fragments

• To separate the DNA fragments by size for sequencing

• To attach fluorescent labels to the DNA fragments

How do fluorescent dideoxynucleotides terminate DNA synthesis in the Sanger method? (NIC)

• By binding to the DNA polymerase

• By cutting the DNA strand

• By incorporating into the DNA strand without a 3' OH group

• By denaturing the DNA double helix

Why are primers designed to be complementary to the target DNA sequence in the Sanger sequencing method?

• To ensure that they bind specifically to the target DNA

• To make them easier to synthesize

• To increase the fluorescence signal

• To allow them to be used in multiple different reactions

What is the main limitation of the Sanger sequencing method? (NIC)

• It can only sequence very short fragments of DNA

• It cannot sequence DNA at all

• It requires a large amount of DNA template

• It is highly inaccurate