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<div class="header">Team</div> | <div class="header">Team</div> | ||
<div class="menu"> | <div class="menu"> | ||
− | <a class="item" href=" https://2016.igem.org/Team:USTC/Team"> Members </a> | + | <a class="item" href="https://2016.igem.org/Team:USTC/Team"> Members </a> |
− | + | <a class="item" href="https://2016.igem.org/Team:USTC/Team/Sponsor"> Sponsor </a> | |
− | + | <a class="item" href="https://2016.igem.org/Team:USTC/Team/AboutUSTC"> About USTC </a> | |
− | + | <a class="item" href="https://2016.igem.org/Team:USTC/Team/Acknowledgements"> Acknowledgements </a> | |
− | + | <a class="item" href="https://2016.igem.org/Team:USTC/Team/Gallery"> Gallery </a> | |
− | + | <a class="item" href="https://2016.igem.org/Team:USTC/Collaborations"> Collaborations </a> | |
</div> | </div> | ||
</div> | </div> | ||
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<div class="header">Team</div> | <div class="header">Team</div> | ||
<div class="menu"> | <div class="menu"> | ||
− | <a class="item" href=" https://2016.igem.org/Team:USTC/Team"> Members </a> | + | <a class="item" href="https://2016.igem.org/Team:USTC/Team"> Members </a> |
− | + | <a class="item" href="https://2016.igem.org/Team:USTC/Team/Sponsor"> Sponsor </a> | |
− | + | <a class="item" href="https://2016.igem.org/Team:USTC/Team/AboutUSTC"> About USTC </a> | |
− | + | <a class="item" href="https://2016.igem.org/Team:USTC/Team/Acknowledgements"> Acknowledgements </a> | |
− | + | <a class="item" href="https://2016.igem.org/Team:USTC/Team/Gallery"> Gallery </a> | |
− | + | <a class="item" href="https://2016.igem.org/Team:USTC/Collaborations"> Collaborations </a> | |
</div> | </div> | ||
</div> | </div> | ||
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</tr> | </tr> | ||
</tbody> | </tbody> | ||
− | <table | + | </table> |
<p>30 cycles(Duration ~ Extend)</p> | <p>30 cycles(Duration ~ Extend)</p> | ||
<p>Agarose gel electrophoresis Results: | <p>Agarose gel electrophoresis Results: |
Latest revision as of 17:55, 19 October 2016
Notebook
Together we stand
Performers
Date: 8.9
Recorder: Kaiyue Ma Ligation of pro+RBS+GFP1-10 & pSB1A3 The concentration of the insert DNA is 60.4 ng/μL. The concentration of the vector DNA is less than 34.6 ng/μL.
material | volume | ||||||
---|---|---|---|---|---|---|---|
Linear vector DNA | 10 μL | ||||||
Insert DNA | 7.8 μL | ||||||
10X buffer | 2 μL | ||||||
T4 DNA ligase | 0.2 μL | ||||||
Total | 20 μL |
sample | 1 | 2 | 3 | 4 | 5 | 6 | 7 |
---|---|---|---|---|---|---|---|
A260/A280 | 1.90 | 1.89 | 1.80 | 1.96 | 1.87 | 1.86 | 1.90 |
A260/A230 | 1.84 | 1.98 | 1.40 | 0.9 | 2.15 | 1.79 | 2.33 |
concentration(ng/μL) | 119.0 | 149.5 | 247.6 | 20.2 | 436.9 | 146.9 | 180.9 |
Material | volume | |||||
---|---|---|---|---|---|---|
dd water | 14.35 μL | |||||
10X buffer | 2 μL | |||||
plasmid (436.9 ng/μL) | 2.3 μL | |||||
FD EcoR I | 1 μL | |||||
FD Pst I | 0.35 μL | |||||
pure love | a lot | |||||
total | 20 μL |
sample | 1 | |||||
---|---|---|---|---|---|---|
A260/A280 | 1.62 | |||||
A260/A230 | 0.12 | |||||
concentration(ng/μL) | 20.6 |
sample | 1 | 2 | 3 | 4 | 5 | |
---|---|---|---|---|---|---|
A260/A280 | 1.86 | 1.88 | 1.83 | 1.86 | 1.86 | |
A260/A230 | 1.38 | 2.21 | 1.76 | 1.94 | 1.95 | |
concentration(ng/μL) | 7.2 | 274.1 | 329.4 | 328.3 | 225.6 |
sample | 1-4 | |||||
---|---|---|---|---|---|---|
Sterilized ddH2O | 32.5 μL | |||||
5×PrimeSTAR Buffer (Mg2+Plus) | 10 μL | |||||
dNTP Mixture (2.5 mM each) | 4 μL | |||||
Template (sfGFP) | 1 μL | |||||
standard-biobrick-p | 1 μL | |||||
standard-biobrick-r | 1 μL | |||||
PrimeSTAR HS DNA Polymerase (2.5 U/μl) | 0.5 μL | |||||
total | 50 μL |
stage | temperature | time | ||||
---|---|---|---|---|---|---|
step 1 | 95 | 5 min | ||||
step 2 | 95 | 10 s | ||||
step 3 | 55 | 15 s | ||||
step 4 | 72 | 12 s | ||||
step 5 | 72 | 10 min | ||||
step 6 | 4 | -- |
stage | temperature | time | ||||
---|---|---|---|---|---|---|
step 1 | 94 | 10 min | ||||
step 2 | 94 | 30 s | ||||
step 3 | 58 | 45 s | ||||
step 4 | 72 | 45 s | ||||
step 5 | 72 | 10 min | ||||
step 6 | 4 | -- |
primer | 1 | 2 | ||||
---|---|---|---|---|---|---|
sequence | 5'-CCGGAATTCGCGGCCGCTTCTAGATGGGCGATTCAAACCAAGGCAACA-3' | 5'-TGCACTGCAGCGGCCGCTACTAGTAATCGTTAACAACTTCGTCATCCACTTCT-3' |
stage | temperature | time | ||||
---|---|---|---|---|---|---|
step 1 | 95 | 7 min | ||||
step 2 | 95 | 10 s | ||||
step 3 | 57 | 15 s | ||||
step 4 | 72 | 50 s | ||||
step 5 | 72 | 10 min | ||||
step 6 | 4 | -- |
sample | 1 | 2 | 3 | 4 | ||
---|---|---|---|---|---|---|
A260/A280 | 1.86 | 1.84 | 1.77 | 1.82 | ||
A260/A230 | 0.76 | 0.36 | 0.31 | 0.25 | ||
concentration(ng/μL) | 137.3 | 120.7 | 60.7 | 25.6 |
number | AD-PYT-4 | BD-PYT-1 | ||||
---|---|---|---|---|---|---|
A260/A280 | 1.74 | 1.64 | ||||
A260/A230 | 1.14 | 1.79 | ||||
concentration(ng/μL) | 45.6 | 166.3 |
primer | 1 | 2 | ||||
---|---|---|---|---|---|---|
sequence | 5'-CCGGAATTCGCGGCCGCTTCTAGATGGGCGATTCAAACCAAGGCAACA-3' | 5'-TGCACTGCAGCGGCCGCTACTAGTAATCGTTAACAACTTCGTCATCCACTTCT-3' |
stage | temperature | time | ||||
---|---|---|---|---|---|---|
step 1 | 95 | 7 min | ||||
step 2 | 95 | 10 s | ||||
step 3 | 57 | 15 s | ||||
step 4 | 72 | 50 s | ||||
step 5 | 72 | 10 min | ||||
step 6 | 4 | -- |
sample | 1 | 2 | 3 | 4 | ||
---|---|---|---|---|---|---|
A260/A280 | 1.70 | 1.86 | 1.85 | 1.85 | ||
A260/A230 | 0.30 | 0.39 | 0.41 | 1.74 | ||
concentration(ng/μL) | 59.1 | 156.5 | 143.2 | 194.5 |
sample | 1,2,3,4 | |||||
---|---|---|---|---|---|---|
Sterilized ddH2O | 9 μL | |||||
Taq PCR Master Mix | 12.5 μL | |||||
Template(bacteria) | 1.5 μL | |||||
Primer 1 | 1 μL | |||||
Primer 2 | 1 μL | |||||
total | 25 μL |
stage | temperature | time | ||||
---|---|---|---|---|---|---|
Pre-Duration | 94 | 4 min | ||||
Duration | 94 | 30 s | ||||
Anneal | 55 | 30 s | ||||
Extend | 72 | 42 s | ||||
Post-Extend | 72 | 10 min | ||||
Final | 4 | -- |
sample | 1-4 | |||||
---|---|---|---|---|---|---|
Sterilized ddH2O | 32.5 μL | |||||
5×PrimeSTAR Buffer (Mg2+Plus) | 10 μL | |||||
dNTP Mixture (2.5 mM each) | 4 μL | |||||
Template (sfGFP) | 1 μL | |||||
standard-sfGFP1-10-p | 1 μL | |||||
standard-sfGFP1-10-r | 1 μL | |||||
PrimeSTAR HS DNA Polymerase (2.5 U/μl) | 0.5 μL | |||||
total | 50 μL |
stage | temperature | time | ||||
---|---|---|---|---|---|---|
step 1 | 95 | 5 min | ||||
step 2 | 95 | 10 s | ||||
step 3 | 55 | 15 s | ||||
step 4 | 72 | 43 s | ||||
step 5 | 72 | 10 min | ||||
step 6 | 4 | -- |
sample | sfGFP1-10-1 | sfGFP1-10-2 | sfGFP1-10-3 | sfGFP1-10-4 | ||
---|---|---|---|---|---|---|
A260/A280 | 1.90 | 1.91 | 1.75 | 1.63 | ||
Concentration(ng/μl) | 140.0 | 143.0 | 33.3 | 34.5 |
Sample | 11μL sfGFP1-10-1 | 11μL sfGFP1-10-2 | 11μL sfGFP1-10-3 | 11μL sfGFP1-10-4 | 15μL pYeT-1 | 15μL pYeT-2 |
---|---|---|---|---|---|---|
nuclease-free water(μL) | 4 | 4 | 4 | 4 | 9 | 9 |
fastdigest buffer(μL) | 2 | 2 | 2 | 2 | 3 | 3 |
PstI(μL) | 1 | 1 | 1 | 1 | 1.5 | 1.5 |
EcoRI(μL) | 1 | 1 | 1 | 1 | 1.5 | 1.5 |
total(μL) | 20 | 20 | 20 | 20 | 30 | 30 |
sample | sfGFP1-10-1(cut) | pYeT |
---|---|---|
A260/A230 | 1.51 | 1.11 |
A260/A280 | 1.92 | 1.85 |
concentration(ng/μL) | 65.5 | 558.8 |
Sample | volume | |
---|---|---|
PYescGAP (cut) | 0.3μL | |
sfGFP1-10 | 1μL | |
nuclease free water | 16.4μL | |
10* T4 DNA Ligase Buffer | 2.0μL | |
T4 DNA Ligase | 0.3μL | |
total | 20μL |
sample | 1 | 2 |
---|---|---|
concentration(ng/μL) | 8.0 | 3.0 |
sample | 1 | 2 |
---|---|---|
A260/A280 | 1.85 | 1.88 |
A260/A230 | 0.58 | 0.18 |
concentration(ng/μL) | 234.9 | 67.8 |
sample | 1 | 2 |
---|---|---|
A260/A280 | 2.01 | 1.85 |
A260/A230 | 0.10 | 0.06 |
concentration(ng/μL) | 8.4 | 9.3 |
sample | 1 | 2 |
---|---|---|
A260/A280 | 29.9 | 1.85 |
A260/A230 | 0.08 | 0.39 |
concentration(ng/μL) | 29.9 | 30.1 |
sample | 1,2,3,4 | |
---|---|---|
Sterilized ddH2O | 16.25 μL | |
5×PrimeSTAR Buffer (Mg2+Plus) | 5 μL | |
dNTP Mixture (2.5 mM each) | 2 μL | |
Template(bacteria) | 1 μL | |
Primer 1 | 0.5 μL | |
Primer 2 | 0.5 μL | |
PrimeSTAR HS DNA Polymerase (2.5 U/μl) | 0.25 μL | |
total | 25.5 μL |
stage | temperature | time |
---|---|---|
Pre-Duration | 95 | 10 min |
Duration | 95 | 10 s |
Anneal | 55 | 15 s |
Extend | 72 | 44 s |
Post-Extend | 72 | 10 min |
Final | 4 | -- |
30 cycles(Duration ~ Extend)
Agarose gel electrophoresis Results: Agarose gel electrophoresis of PCR product (from left to right: marker,control,sample 1,2,3,4)
Failed
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