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− | <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> | |
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<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> | |
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<p><strong>PCR of sfGFP11</strong> | <p><strong>PCR of sfGFP11</strong> | ||
<strong>Recorder: Xuefeng Meng</strong> | <strong>Recorder: Xuefeng Meng</strong> |
Revision as of 18:24, 19 October 2016
Notebook
Together we stand
Performers
DATE:8.24
Recorder: Yu Xie Plasmid Extraction for sfGFP11
Recorder: Yu Xie PCR of sup35
Procedure:
1.Prepare 8 PCR tubes and sequentially add:
sample | 1,2,3,4,5,6,7,8 | |||
---|---|---|---|---|
Sterilized ddH2O | 9 μL | |||
2×HiFi-PCR Master | 12.5 μL | |||
Template | 1.5 μL | |||
Primer 1 | 1 μL | |||
Primer 2 | 1 μL | |||
total | 25 μL |
stage | temperature | time | ||
---|---|---|---|---|
Pre-Duration | 95℃ | 10 min | ||
Duration | 95℃ | 1 min | ||
Anneal | 58℃ | 1 min | ||
Extend | 72℃ | 1 min 50 s | ||
Post-Extend | 72℃ | 10 min | ||
Final | 4℃ | -- |
sample | 1,2,3,4,5,6,7,8 | |||
---|---|---|---|---|
Sterilized ddH2O | 9 μL | |||
2×HiFi-PCR Master | 12.5 μL | |||
Template | 1.5 μL | |||
Primer 1 | 1 μL | |||
Primer 2 | 1 μL | |||
total | 25 μL |
stage | temperature | time | ||
---|---|---|---|---|
Pre-Duration | 95℃ | 10 min | ||
Duration | 95℃ | 1 min | ||
Anneal | 58℃ | 1 min | ||
Extend | 72℃ | 1 min 50s | ||
Post-Extend | 72℃ | 10 min | ||
Final | 4℃ | -- |
stage | temperature | time | ||
---|---|---|---|---|
step 1 | 95℃ | 10 min | ||
step 2 | 95℃ | 30 s | ||
step 3 | 58℃ | 45 s | ||
step 4 | 72℃ | 45 s | ||
step 5 | 72℃ | 10 min | ||
step 6 | 4℃ | -- |
sample | YBE8241001 | YBE8241002 | YBE8241003 | YBE8241004 |
---|---|---|---|---|
A260/A280 | 1.88 | 1.86 | 1.89 | 1.88 |
A260/A230 | 2.27 | 1.92 | 2.22 | 2.22 |
concentration(ng/μL) | 203.1 | 174.7 | 181.3 | 177.8 |
stage | temperature | time | |
---|---|---|---|
step 1 | 95℃ | 10 min | |
step 2 | 95℃ | 30 s | |
step 3 | 58℃ | 45 s | |
step 4 | 72℃ | 45 s | |
step 5 | 72℃ | 10 min | |
step 6 | 4℃ | -- |
sample | 1,2,3,4,5,6,7,8 | ||
---|---|---|---|
Sterilized ddH2O | 9 μL | ||
2×HiFi-PCR Master | 12.5 μL | ||
Template | 1.5 μL | ||
Primer 1 | 1 μL | ||
Primer 2 | 1 μL | ||
total | 25 μL |
stage | temperature | time | |
---|---|---|---|
Pre-Duration | 95℃ | 10 min | |
Duration | 95℃ | 1 min | |
Anneal | 58℃ | 1 min | |
Extend | 72℃ | 1 min 50 s | |
Post-Extend | 72℃ | 10 min | |
Final | 4℃ | -- |
stage | temperature | time | |
---|---|---|---|
step 1 | 95℃ | 10 min | |
step 2 | 95℃ | 30 s | |
step 3 | 58℃ | 45 s | |
step 4 | 72℃ | 45 s | |
step 5 | 72℃ | 10 min | |
step 6 | 4℃ | -- |
sample | 1 | 2 | 3 |
---|---|---|---|
A260/A280 | 2.02 | 1.90 | 1.91 |
A260/A230 | 1.19 | 1.07 | 1.31 |
concentration(ng/μL) | 17.0 | 15.5 | 27.9 |
PCR of sfGFP11 Recorder: Xuefeng Meng results