Sanger Sequencing

What are the ddNTPs missing?

• A 3rd OH

• A Carbon atom

• A phosphate group

• The 4th OH

Why is modern Sanger sequencing possible in 1 tube?

• Use of radioisotopes

• Colored gel is used now

• Fluorescent tagging

• Genic modification

• Scientists don't want to waste 4 tubes any more

Interpret this gel to give the base sequence of the replicated strand (NOT the template strand) [without spaces]

Tags

NGSS.HS-LS1-1

What happens when a DNA polymerase uses a ddNTP?

• Strand synthesis proceeds as usual

• The strand becomes heavier

• The whole DNA molecule breaks down

• Strand synthesis is terminated

Full form of ddNTP

• Di deoxy nucleotide triphosphate

• Demi deoxy nucleotide triphosphate

• Deca deoxy nucleotide triphosphate

• Di deoxy nucleotide tetraphosphate

Interpret the FINAL template DNA base sequence

• GGCCTGTCTTCAGAAA

• GCTAGCTAGGTCGATC

• AAAGACTTCTGTCCGG

• CCGGACAGAAGTCTTT

Tags

NGSS.HS-LS1-1