Steps of DNA Sequencing

Question 1

First step

Answer 1

Isolate DNA

Question 2

Second step

Answer 2

1st PCR (in thermal cycler)
Amplify the pathogen DNA only (making copies)
TUBE CONTAINS: DNA polymerase, dNTP (nucleotides), DNA primers, pathogen DNA, Sue’s DNA
30 cycles of
Denaturing, annealing, extension

Question 3

Third step

Answer 3

Cycle sequencing
2nd PCR
tag DNA fragments with fluorescent nucleotides
3 steps are the same except for the extension (when DNA polymerase adds a tagged nucleotide (ddNTP) the reaction steps and no more nucleotides are added)