DNA synthesis involves a reaction between the 3'-hydroxyl group of the primer dNTP and the phosphate group of an incoming dNTP. The Sanger method uses ddNTPs, which lack the 3'-hydroxyl group, to halt DNA synthesis at a particular nucleotide. In the sequencing reaction, DNA synthesis is carried out with a mixture of dNTPs and a ddNTP to extend a radiolabeled primer. A different ddNTP is used in each reaction. The products are analyzed by autoradiography to determine the nucleotide sequence.
Sanger Sequencing - Exercise
Detailed view of Sanger Sequencing reactions...
Congratulations, you successfully terminated the DNA synthesis with a ddNTP.
You failed to end the DNA synthesis with a ddNTP, press the try again button.
Sanger Sequencing - Introduction
In this excercise, we will be sequencing the BRCA-II gene. Your job at the end of the exercise will be to write out the nucleotide sequence.
Sanger Sequencing - Simulation
Close-up view of Sanger sequencing reaction...
When the sequencing reaction completes, the products will be moved into a capillary gel and read by a laser. In order to see this, please scroll the page down. Once the chromatogram is completed, please enter the sequence of the BRCA-II gene.
The number of nucleotides you entered does not match the chromatogram. Please re-enter your answer and click "Submit DNA" again.Your answer contains non-nucleotide letters. Please re-enter your answer and click "Submit DNA" again.
RNA Processing - Simulation Results
Your Answer:
Correct Answer:
NOTE: This is not the actual sequence of the BRCA-II gene, but rather just a random sequence created for this exercise.
Results:
Congratulations! You figured out the sequence based on the chromatogram. If you would like to try the exercise again with a new dna sequence, please click the button below.
Results:
Sorry, you selected the wrong nucleotides. If you would like to try the exercise again with a new dna sequence, please click the button below.
