{{ construct.name }} | http://django.gibthon.org/gibthon/{{ construct.id }}/{{ construct.name }}/ |
{{ construct.description|linebreaks }}
Fragments: {{ construct.fragments.all.count }}
Features: {{construct.feature_count }}
Total lenght: {{ construct.length }} bp |
![]() Generated by Gibthon Construct Designer |
Gibson Assembly Protocol:{{ construct.name }} |
Assembly Mix | Volume |
---|---|
Gibson Master Mix | 15 μl |
{{ f.name }} (extended) | {{ each|floatformat:1 }} μl |
Total | 20 μl |
Gibson Master Mix | Volume |
Taq ligase (40u/µl) | 50 μl |
5x isothermal buffer | 100 μl |
T5 exonuclease (1u/µl) | 2 μl |
Phusion polymerase (2u/µl) | 6.25 μl |
Nuclease-free water | 216.75 μl |
Total | 375 μl |
5x isothermal buffer | Volume |
25% PEG-8000 | 0.75g |
500 mM Tris-HCl pH 7.5 | 1500 μl |
50mM MgCl2 | 75 μl |
50mM DTT | 150 μl |
1mM dATP | 30 μl |
1mM dTTP | 30 μl |
1mM dCTP | 30 μl |
1mM dGTP | 30 μl |
5mM NAD | 300 μl |
Nuclease-free water | ... |
Total | 3000 μl |
{{ cf.fragment.name }}:{{ construct.name }} | Primer datasheet |
Section | Full | Anneal | |
---|---|---|---|
Length | {{ primer.length }} bp | {{ primer.stick.length }} bp | |
Tm | {{ primer.tm|floatformat }}°C | {{ primer.stick.tm|floatformat }}°C | |
Sequence | {{ primer.seq_pretty }} |
Section | Full | Anneal | |
---|---|---|---|
Length | {{ primer.length }} bp | {{ primer.stick.length }} bp | |
Tm | {{ primer.tm|floatformat }}°C | {{ primer.stick.tm|floatformat }}°C | |
Sequence | {{ primer.seq_pretty }} |
PCR Mix | Volume |
---|---|
Buffer | {{ construct.pcrsettings.buffer_v|floatformat:1 }} μl |
dNTPs | {{ construct.pcrsettings.dntp_v|floatformat:1 }} μl |
{{ cf.primer_top.name }} | {{ cf.primer_top.vol|floatformat:1 }} μl |
{{ cf.primer_bottom.name }} | {{ cf.primer_bottom.vol|floatformat:1 }} μl |
{{ cf.fragment.name }} | {{ cf.vol|floatformat:1 }} μl |
Enzyme | {{ construct.pcrsettings.enzyme_v|floatformat:1 }} μl |
Water | {{ cf.water_v|floatformat:1 }} μl |
Total Volume | {{ construct.pcrsettings.total_v|floatformat:1 }} μl |
PCR Instructions | Temperature | Time |
---|---|---|
Initial Melting | 98°C | 30s |
cycle 30 times | ||
Melting | 98°C | 30s |
Annealing | {{ cf.tm|floatformat:0 }}°C | 30s |
Elongation | 72°C | {{ cf.time|floatformat:0 }}s |
end cycle | ||
Final Elongation | 98°C | 7m 30s |
Final Hold | 98°C | ∞ |