Where and for what reason (s) are you likely to observe the letter "N" in your Sanger sequence reads? Select all that apply. Near the end of a read (which is typically about 500-700 bp in length); DNA polymerase will nearly always incorporate a ddNTP into a read when it reaches this length. Anywhere a mutation leads to incorporation of a translation stop codon in the DNA; DNA polymerase is unable to incorporate ddNTPs into this sequence Anywhere the candidate differs from the reference sequence; any deviation from the reference sequence will inhibit the ability of the sequence analyzer to correctly interpret the base In the first 25-100 base pairs of the read; ddNTPs are unlikely to be incorporated in this region because they are greatly outnumbered by dNTPs.
Near the end of a read (which is typically about 500-700 bp in length); DNA polymerase will nearly always incorporate a ddNTP into a read when it reaches this length.
Anywhere a mutation leads to incorporation of a translation stop codon in the DNA; DNA polymerase is unable to incorporate ddNTPs into this sequence
Anywhere the candidate differs from the reference sequence; any deviation from the reference sequence will inhibit the ability of the sequence analyzer to correctly interpret the base
In the first 25-100 base pairs of the read; ddNTPs are unlikely to be incorporated in this region because they are greatly outnumbered by dNTPs.
Sanger sequencing, also known as chain-termination sequencing, is a method for determining the sequence of nucleotide bases in a DNA molecule. The method uses a DNA polymerase to synthesize a complementary strand of DNA using a single-stranded template DNA, which is annealed with a specific primer. In each cycle of the reaction, a dideoxynucleotide (ddNTP) is incorporated into the growing DNA strand, which terminates chain elongation. The resulting DNA fragments are separated by size using gel electrophoresis, and the sequence is read by determining the order of the terminating nucleotides. The method is named after its inventor, Frederick Sanger, and it was the first DNA sequencing technique to be widely used.
Step by step
Solved in 2 steps
