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MGG exam 3Wha – Questions with Answers
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MGG exam 3Wha – Questions with Answers
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MGG exam 3Wha – Questions withAnswers
What are the three basic steps of 454 sequencing - - start with complex mix
of DNA
emulsion PCR
sequencing
- Describe the first step of 454 sequencing - - DNA fragments of PCR
amplified DNA
adapter DNA us ligated to each end of DNA
- describe the adaptor DNA in 454 sequencing - - Adaptor B binds with DNA
primer attached to bead
Adaptor A binds to free primer after replication
- Describe emulsion PCR in 454 sequencing - - B adapter binds to primer on
B and PCR occurs
free primer is added and binds to adapter A and source DNA is amplified
3:1 ratio of beads to DNA
water mixed with oil for millions of reaction chambers
- Describe sequencing in 454 - - break emulsion and recover beads
add them to pico wells
fill with lightning beads
add dNTP one at a time
if incorporated, a flash of light occurs
height of peak indicates single or multiple bases in a row
- describe the enzymes in 454 sequencing - - sulfurylase- ATP creation from
APS and PP
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