Draw a workflow of the original Sanger Sequencing Method
In automated DNA sequencing reaction by the Sanger method, which of the following is NOT a component of the reaction? A) fluorescently-labeled deoxynucleotides B) DNA polymerase C) a DNA primer D) the DNA to be sequenced E) none of the above (i.e. all the above are required in the reaction)
The data below were obtained as the result of the Sanger method of DNA sequencing. Determine the DNA sequence. making sure to indicate 5 and 3 ends, and add the sequence for the complementary strand
O The Sanger method of DNA sequencing requires which of the following to be in the reaction tube? Choose one: O A. template DNA O B. deoxyribonucleotides O C. dideoxyribonucleotides O D. A, B, and C are correct. O E. Only A and B are correct. O F. Only A and Care correct. O O O
ddATP ddCTP ddGTP ddTTP The autoradiogram shown was obtained from a DNA sequencing method called Sanger sequencing or dideoxy sequencing. The arrow shows the direction of migration of the DNA samples during electrophoresis. Determine the sequence of the DNA template strand, in the 5' to 3' direction, from this data. 5' – CGTCAATTTAG
Compare Sanger sequencing and NGS 1. Sanger Sequencing: Amount of DNA (high/low)? Sequence mixed DNA (Yes/No)? Efficiency (high/low)? 2. NGS Amount of DNA (high/low)? Sequence mixed DNA (Yes/No)? Efficiency (high/low)?
Sanger sequencing or dideoxy sequencing of DNA causes termination of replication. Why? 18.
Provide four ways that cycle sequencing is an improvement over conventional Sanger sequencing.
The Sanger method of DNA sequencing requires what type of dNTP modification to generate truncated DNA products? How are individual dNTPs identified? How are the different sizes of DNA separated from each other?
The Sanger method of DNA sequencing uses which chain-terminating ingredient to separate fragments differing by a single nucleotide a. Magnesium b. Heat c. ddNTPs d. restriction enzymes e. none of the above
Q1) List the limitations and challenges of dideoxy termination method of DNA sequencing? Q2) In the context of mutation detection, describe the differences between first- and second-generation sequencing? Q3) 6-year-old child presenting with sensorineural deafness, sequencing of MYO15A gene by Sanger method was performed and showed the following chromatogram: A- Describe the principle of Sanger sequencing method? B- Describe the DNA change: position, type of variant and genotype?