Completing the Lagging Strand [Diagram]
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Completing the Lagging Strand [Diagram]
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Chain Termination PCR
-- also called Sanger sequencing
-- used to determine the nucleotide sequence of DNA
-- manual or automated
-- PCR occurs first
-- DNA is denatured to break the double-strand into two single strands
-- primers anneal to each strand
-- they extend to create two double strands
-- in Sanger sequencing / chain termination, ddNTPs are added along with dNTPs
-- ddNTPs can be added randomly as the chain extends
-- they stop the chain
-- this is because they can’t form phosphodiester bonds like dNTPs can
-- next, the products are separated by size
-- this is done with gel electrophoresis
-- DNA polymerase works in the 5′ to 3′ direction
-- each terminal ddNTP will correspond to a specific nucleotide in the original sequence
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