DNA sequincing Flashcards
who invented the sanger dideoxy method
Frederick sanger in 1977
what is the sanger dideoxy method
DNA was treated to become a single strand (by using heat)
A short, single-stranded, radioactively labeled primer was added to the end of the strand
This will cause the other side to begin synthesizing
This in return will allow the template strand to be decoded
DNA polymerase was added to build the strand
Regular nucleotides were added along with special radioactively labeled dideoxy analogs
The dideoxy analog is any nucleotide whose deoxyribose sugar is missing the OH group on carbon3’
This OH is needed to attach to the phosphate group on the next nucleotide (STOPS THE BUILDING)
Radioactive components glow
This will determine one letter at a time meaning the process must be repeated constantly until every letter in the section by random can be identified
why were radioactively labeled primers discontinued and what were they changed to?
- radiation was too toxic and too dangerous
- changed to florescent tagged dideoxy angelus (easier to see as well as safer)
