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Phusion: Difference between revisions
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→Thermocycling conditions
(→Thermocycling conditions: comment on Tm calculation method) |
Jarle Pahr (talk | contribs) |
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#15-30 s/kb extension time. | #15-30 s/kb extension time. | ||
#98C for denaturation. | #98C for denaturation. | ||
#Anneal at 3C above the Tm. | #Anneal at 3C above the lowest Tm if the primers are longer then 20nt, esle at the Tm. | ||
#*Note: Tm should be calculated with nearest neighbor method see: [https://www.finnzymes.fi/tm_determination.html Finnzyme Tm calculator] | #*Note: Tm should be calculated with nearest neighbor method see: [https://www.finnzymes.fi/tm_determination.html Finnzyme Tm calculator] | ||
#Extend at 72C. | #Extend at 72C. | ||
