Part:BBa_K187202:Experience
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Applications of BBa_K187202
Source of part:
Oligonucleotides were synthesized. Primers were tested using MG1655 E.coli genomic DNA as template.
Design considerations:
This primer produced a product of the predicted size using the following reaction conditions:
Water: 17.05uL
10x pfu buffer: 2.5uL
dNTPs (2mM): 2.5uL
DMSO: 1.2uL
MG1655 Genomic DNA: 0.5uL
Forward primer (10uM): 0.5uL
Reverse primer (10uM): 0.5uL
Pfu polymerase: 0.25uL
Total reaction volume: 25uL
Thermocycling conditions:
95oC, 3min
95oC, 30s
56oC, 30s
72oC, 3min
29 cycles to step 2
72oC 2min
All Biobytes essential gene primers were produced using Vector NTI. Each primer's thermodynamic properties were examined using Vector NTI's hairpin and dimerzation programs. Primers were optimized to fit as many of the following criteria as possible:
Find the shortest possible sequence, reducing the cost to produce the primer. Produce the highest score value possible. Produce the closest Tm's possible Produce hairpins with dG values >-5 Produce dimers with dG values >-10
User Reviews
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