Team:Kyoto/Notebook
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<p><a href="https://2014.igem.org/Team:Kyoto/noteame">アメフラシはこちら</a></p> | <p><a href="https://2014.igem.org/Team:Kyoto/noteame">アメフラシはこちら</a></p> | ||
+ | |||
+ | |||
+ | |||
+ | |||
+ | |||
+ | <h4>Liquid Culture</h4> | ||
+ | <span class=“kyoto-author">名前</span> | ||
+ | <table> | ||
+ | <tr><th>Sample</th><th>Medium</th></tr> | ||
+ | <tr><td>サンプル名</td><td>培地</td></tr> | ||
+ | </table> | ||
+ | |||
+ | |||
+ | <h4>Gel Extraction</h4> | ||
+ | <span class=“kyoto-author">名前</span> | ||
+ | <table> | ||
+ | <tr><th>Lane</th><th>Sample</th></tr> | ||
+ | <tr><td>レーン番号</td><td>サンプル名</td></tr> | ||
+ | </table> | ||
+ | |||
+ | |||
+ | <h4>Electrophoresis</h4> | ||
+ | <span class=“kyoto-author">名前</span> | ||
+ | <table> | ||
+ | <tr><th>Lane</th><th>Sample</th><th>Ensyme1</th><th>Ensyme2</th></tr> | ||
+ | <tr><td>レーン番号</td><td>サンプル名</td><td>制限酵素1</td><td>制限酵素2</td></tr> | ||
+ | </table> | ||
+ | |||
+ | |||
+ | <h4>Miniprep/DNA Purification</h4> | ||
+ | <span class=“kyoto-author">名前</span> | ||
+ | <table> | ||
+ | <tr><th>Sample</th><th>Concentration/(µg/mL)</th><th>260/280</th><th>260/230</th></tr> | ||
+ | <tr><td>サンプル名</td><td>(Concentration)</td><td>(260/280)</td><td>(260/280)</td></tr> | ||
+ | </table> | ||
+ | |||
+ | |||
+ | <h4>Restriction Enzyme Digestion</h4> | ||
+ | <span class=“kyoto-author">名前</span> | ||
+ | <table> | ||
+ | <tr><th colspan=2>Sample/(µL)</th><th>EcoR1/(µL)</th><th>Xba1/(µL)</th><th>Spe1/(µL)</th><th>Pst1/(µL)</th><th colspan=2>Buffer/(µL)</th><th>BSA/(µL)</th><th>MilliQ/(µL)</th><th>Total/(µL)</tr> | ||
+ | <tr><td>サンプル名</td><td>(サンプル)</td><td>(E)</td><td>(X)</td><td>(S)</td><td>(P)</td><td>(HorM)<td>(Buffer)</td></td><td>(BSA)</td><td>(MilliQ)</td><td>(Total)</td></tr> | ||
+ | </table> | ||
+ | |||
+ | |||
+ | <h4>Ligation</h4> | ||
+ | <span class=“kyoto-author">名前</span> | ||
+ | <table> | ||
+ | <tr><th colspan=2>Vector/(µL)</th><th colspan=2>Insert/(µL)</th><th colspan=2>Buffer/(µL)</th><th colspan=2>Ligase/(µL)</th><th>MilliQ/(µL)</th><th>Total/(µL)</tr> | ||
+ | <tr><td>ベクター名</td><td>(ベクター)</td><td>インサート名</td><td>(インサート)</td><td>(HorM)</td><td>(Buffer)</td><td>Ligase名</td><td>(Ligase)</td><td>(MilliQ)</td><td>(Total)</td></tr> | ||
+ | </table> | ||
+ | |||
+ | |||
+ | <h4>Transformation</h4> | ||
+ | <span class=“kyoto-author">名前</span> | ||
+ | <table> | ||
+ | <tr><th colspan=2>Sample/(µL)</th><th colspan=2>CompetentCells/(µL)</th><th>Total/(µL)</th><th>Medium</th></tr> | ||
+ | <tr><td>サンプル名</td><td>(サンプル)</td><td>コンピ名</td><td>(コンピ)</td><td>(Total)</td><td>培地</td></tr> | ||
+ | </table> | ||
+ | |||
+ | |||
+ | <h4>Dephosphorylation</h4> | ||
+ | <span class=“kyoto-author">名前</span> | ||
+ | <table> | ||
+ | <tr><th colspan=2>Sample/(µL)</th></tr> | ||
+ | <tr><td>サンプル名</td><td>(サンプル)</td></tr> | ||
+ | </table> | ||
+ | |||
+ | |||
+ | |||
+ | |||
+ | |||
+ | |||
+ | |||
+ | |||
+ | |||
<h2>7/15</h2> | <h2>7/15</h2> | ||
<h3>transformation</h3> | <h3>transformation</h3> |
Revision as of 08:14, 23 September 2014
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Home | Team | Official Team Profile | Project | Parts | Modeling | Notebook | Safety | Attributions |
Notebook
Calendar
February | ||||||
---|---|---|---|---|---|---|
1 | ||||||
2 | 3 | 4 | 5 | 6 | 7 | 8 |
9 | 10 | 11 | 12 | 13 | 14 | 15 |
16 | 17 | 18 | 19 | 20 | 21 | 22 |
23 | 24 | 25 | 26 | 27 | 28 |
March | ||||||
---|---|---|---|---|---|---|
1 | ||||||
2 | 3 | 4 | 5 | 6 | 7 | 8 |
9 | 10 | 11 | 12 | 13 | 14 | 15 |
16 | 17 | 18 | 19 | 20 | 21 | 22 |
23 | 24 | 25 | 26 | 27 | 28 | 29 |
30 | 31 |
July | ||||||
---|---|---|---|---|---|---|
1 | 2 | 3 | 4 | 5 | ||
6 | 7 | 8 | 9 | 10 | 11 | 12 |
13 | 14 | 15 | 16 | 17 | 18 | 19 |
20 | 21 | 22 | 23 | 24 | 25 | 26 |
27 | 28 | 29 | 30 | 31 |
August | ||||||
---|---|---|---|---|---|---|
1 | 2 | |||||
3 | 4 | 5 | 6 | 7 | 8 | 9 |
10 | 11 | 12 | 13 | 14 | 15 | 16 |
17 | 18 | 19 | 20 | 21 | 22 | 23 |
24 | 25 | 26 | 27 | 28 | 29 | 30 |
31 |
September | ||||||
---|---|---|---|---|---|---|
1 | 2 | 3 | 4 | 5 | 6 | |
7 | 8 | 9 | 10 | 11 | 12 | 13 |
14 | 15 | 16 | 17 | 18 | 19 | 20 |
21 | 22 | 23 | 24 | 25 | 26 | 27 |
28 | 29 | 30 |
Octorber | ||||||
---|---|---|---|---|---|---|
1 | 2 | 3 | 4 | |||
5 | 6 | 7 | 8 | 9 | 10 | 11 |
12 | 13 | 14 | 15 | 16 | 17 | 18 |
19 | 20 | 21 | 22 | 23 | 24 | 25 |
26 | 27 | 28 | 29 | 30 | 31 |
Liquid Culture
Sample | Medium |
---|---|
サンプル名 | 培地 |
Gel Extraction
Lane | Sample |
---|---|
レーン番号 | サンプル名 |
Electrophoresis
Lane | Sample | Ensyme1 | Ensyme2 |
---|---|---|---|
レーン番号 | サンプル名 | 制限酵素1 | 制限酵素2 |
Miniprep/DNA Purification
Sample | Concentration/(µg/mL) | 260/280 | 260/230 |
---|---|---|---|
サンプル名 | (Concentration) | (260/280) | (260/280) |
Restriction Enzyme Digestion
Sample/(µL) | EcoR1/(µL) | Xba1/(µL) | Spe1/(µL) | Pst1/(µL) | Buffer/(µL) | BSA/(µL) | MilliQ/(µL) | Total/(µL) | ||
---|---|---|---|---|---|---|---|---|---|---|
サンプル名 | (サンプル) | (E) | (X) | (S) | (P) | (HorM) | (Buffer) | (BSA) | (MilliQ) | (Total) |
Ligation
Vector/(µL) | Insert/(µL) | Buffer/(µL) | Ligase/(µL) | MilliQ/(µL) | Total/(µL) | ||||
---|---|---|---|---|---|---|---|---|---|
ベクター名 | (ベクター) | インサート名 | (インサート) | (HorM) | (Buffer) | Ligase名 | (Ligase) | (MilliQ) | (Total) |
Transformation
Sample/(µL) | CompetentCells/(µL) | Total/(µL) | Medium | ||
---|---|---|---|---|---|
サンプル名 | (サンプル) | コンピ名 | (コンピ) | (Total) | 培地 |
Dephosphorylation
Sample/(µL) | |
---|---|
サンプル名 | (サンプル) |
7/15
transformation
Name | Sample/(µl) | Competent Cells/(µl) | Total/(µl) | Medium |
---|---|---|---|---|
BBa_J23101(Pcon) | 2 | 20(2/23) | 22 | LB(CP) |
BBa_B0032(RBS) | 2 | 20(2/23) | 22 | LB(CP) |
7/16
transformation
Name | Sample/(µl) | Competent Cells/(µl) | Total/(µl) | Medium |
---|---|---|---|---|
BBa_B0030(1) | 2 | 20(2/23) | 22 | LB(CP) |
BBa_B0030(2) | 2 | 20(bought) | 22 | LB(CP) |
BBa_B0032(1) | 2 | 20(2/23) | 22 | LB(CP) |
BBa_B0032(2) | 2 | 20(bought) | 22 | LB(CP) |
7/17
Colony PCR
Name |
---|
7/16 BBa_B0030(1) Colony_1 |
7/16 BBa_B0030(1) Colony_2 |
7/16 BBa_B0030(1) Colony_3 |
7/16 BBa_B0030(2) Colony_1 |
7/16 BBa_B0030(2) Colony_2 |
7/16 BBa_B0030(2) Colony_3 |
7/16 BBa_B0032(1) Colony_1 |
7/16 BBa_B0032(1) Colony_2 |
7/16 BBa_B0032(1) Colony_3 |
7/16 BBa_B0032(2) Colony_1 |
7/16 BBa_B0032(2) Colony_2 |
7/16 BBa_B0032(2) Colony_3 |
Negative control |
Liquid Culture
Name | medium |
---|---|
7/16 BBa_B0030(1) Colony_1 | Plusgrow medium(CP) |
7/16 BBa_B0030(1) Colony_2 | Plusgrow medium(CP) |
7/16 BBa_B0030(1) Colony_3 | Plusgrow medium(CP) |
7/16 BBa_B0030(2) Colony_1 | Plusgrow medium(CP) |
7/16 BBa_B0030(2) Colony_2 | Plusgrow medium(CP) |
7/16 BBa_B0030(2) Colony_3 | Plusgrow medium(CP) |
7/16 BBa_B0032(1) Colony_1 | Plusgrow medium(CP) |
7/16 BBa_B0032(1) Colony_2 | Plusgrow medium(CP) |
7/16 BBa_B0032(1) Colony_3 | Plusgrow medium(CP) |
7/16 BBa_B0032(2) Colony_1 | Plusgrow medium(CP) |
7/16 BBa_B0032(2) Colony_2 | Plusgrow medium(CP) |
7/16 BBa_B0032(2) Colony_3 | Plusgrow medium(CP) |
7/18
Miniprep
DNA | Concentration/(µg/ml) | 260/280 | 260/230 |
---|---|---|---|
7/16 BBa_B0030(1) Colony_1 | 229 | 1.96 | 1.40 |
7/16 BBa_B0030(1) Colony_2 | 209 | 1.95 | 1.50 |
7/16 BBa_B0030(1) Colony_3 | 108 | 2.12 | 2.12 |
7/16 BBa_B0030(2) Colony_1 | - | - | - |
7/16 BBa_B0030(2) Colony_2 | 174 | 2.02 | 1.56 |
7/16 BBa_B0030(2) Colony_3 | 158 | 2.01 | 1.68 |
7/16 BBa_B0032(1) Colony_1 | 179 | 2.00 | 2.06 |
7/16 BBa_B0032(1) Colony_2 | - | - | - |
7/16 BBa_B0032(1) Colony_3 | 221 | 1.90 | 1.37 |
7/16 BBa_B0032(2) Colony_1 | 225 | 1.89 | 1.37 |
7/16 BBa_B0032(2) Colony_2 | 270 | 1.98 | 1.63 |
7/16 BBa_B0032(2) Colony_3 | 194 | 1.89 | 1.33 |
7/30
Restriction Enzyme Digestion
Sample | DNA/(µl) | EcoR1/(µl) | Xba1/(µl) | Spe1/(µl) | Pst1/(µl) | Buffur/(µl) | BSA/(µl) | MilliQ/(µl) | TOTAL/(µl) |
---|---|---|---|---|---|---|---|---|---|
7/16 BBa_B0030(2) Colony_1 | 1 | 0.3 | 0 | 0 | 0 | 1(H) | 0 | 7.7 | 10 |
7/16 BBa_B0030(2) Colony_1 | 1 | 0 | 0.3 | 0 | 0 | 1(M) | 1 | 6.7 | 10 |
7/16 BBa_B0030(2) Colony_1 | 1 | 0 | 0 | 0.3 | 0 | 1(M) | 0 | 7.7 | 10 |
7/16 BBa_B0030(2) Colony_1 | 1 | 0 | 0 | 0 | 0.3 | 1(H) | 0 | 7.7 | 10 |
Control | 1 | 0 | 0 | 0 | 0 | 1(M) | 0 | 8 | 10 |
Electrophoresis for confirmation
Lane | Restriction Enzyme Digestion Products |
---|---|
1 | Product1 |
2 | Product2 |
3 | Product3 |
4 | Product4 |
5 | Product5 |
6 | 1kbp ladder |
7/31
Transformation
Name | Sample/(µl) | Competent Cells/(µl) | Total/(µl) | Medium |
---|---|---|---|---|
BBa_E0240(RBS-GFP-DT) | 2 | 20 | 22 | LB(CP) |
J23100(Pcon) | 2 | 20 | 22 | LB(AMP) |
Liquid Culture
Name | medium |
---|---|
7/31 J23100(Pcon) Colony_1 | Plusgrow medium(AMP) |
7/31 J23100(Pcon) Colony_2 | Plusgrow medium(AMP) |
7/31 J23100(Pcon) Colony_3 | Plusgrow medium(AMP) |
7/31 J23100(Pcon) Colony_4 | Plusgrow medium(AMP) |
7/31 J23100(Pcon) Colony_5 | Plusgrow medium(AMP) |
7/31 J23100(Pcon) Colony_6 | Plusgrow medium(AMP) |
Restriction Enzyme Digestion
7/18 RBS Sample | DNA/(µl) | EcoR1/(µl) | Spe1/(µl) | Pst1/(µl) | Xba1/(µl) | Buffur/(µl) | BSA/(µl) | MilliQ/(µl) | TOTAL/(µl) |
---|---|---|---|---|---|---|---|---|---|
BBa_B0032 | 5 | 0.3 | 0 | 0 | 0 | 1(H) | 0 | 3.7 | 10 |
BBa_B0032 | 5 | 0 | 0.3 | 0 | 0 | 1(M) | 0 | 3.7 | 10 |
BBa_B0032 | 5 | 0 | 0 | 0.3 | 0 | 1(H) | 0 | 3.7 | 10 |
BBa_B0032 | 5 | 0 | 0 | 0 | 0.3 | 1(M) | 1 | 2.7 | 10 |
Control | 5 | 0 | 0 | 0 | 0 | 1(M) | 0 | 4 | 10 |
8/1
Electrophoresis for confirmation
Lane | Restriction Enzyme Digestion Products |
---|---|
1 | 1kb ladder |
2 | Product1 |
3 | Product2 Error |
4 | - |
5 | - |
6 | Product3 |
7 | Product4 |
8 | Product5 |
8/1
PCR
BBa_E0240(RBS-GFP-DT) | Volume |
---|---|
10×buffur | 5μL |
2mMdNTPs | 5μL |
25mM MgSO4 | 2μL |
VF Primer | 1.5μL |
VR Primer | 1.5μL |
template | 0.5μL |
KOD-plus | 1μL |
MilliQ | 33.5μL |
TOTAL | 50μL |
Steps | Temparature | Time | cycle |
---|---|---|---|
Pre Denature | 94℃ | 2min | 1 time |
Denature | 94℃ | 15sec | 30 cycle |
Annealing | 55℃ | 30sec | 30 cycle |
Extension | 68℃ | 1min | 30 cycle |
DNA | Concentration/(µg/ml) | 260/280 | 260/230 |
---|---|---|---|
Product | 80 | 1.91 | 1.78 |
8/2
Miniprep
DNA | Concentration/(µg/ml) | 260/280 | 260/230 |
---|---|---|---|
7/31 J23100(Pcon) Colony_1 | 145 | 2.11 | 3.09 |
7/31 J23100(Pcon) Colony_2 | - | - | - |
7/31 J23100(Pcon) Colony_3 | 160 | 2.01 | 3.13 |
7/31 J23100(Pcon) Colony_4 | 140 | 1.87 | 2.70 |
7/31 J23100(Pcon) Colony_5 | 105 | 2.39 | 5.90 |
7/31 J23100(Pcon) Colony_6 | 180 | 1.90 | 1.37 |
8/20
Transformation
Name | Sample/(µl) | Competent Cells/(µl) | Total/(µl) | Medium |
---|---|---|---|---|
7/31 BBa_E0240(RBS-GFD-DT) | 2 | 20 | 22 | LB(CP) |
BBa_I732095 | 2 | 20 | 22 | LB(CP) |
8/21
Colony PCR
Name |
---|
8/20 BBa_I732095 Colony_1 |
8/20 BBa_I732095 Colony_2 |
8/20 BBa_I732095 Colony_3 |
Negative control |
Electrophoresis for confirmation
Lane | PCR Product |
---|---|
1 | 1kb ladder |
2 | 8/21 BBa_I732095 Colony_1 |
3 | 8/21 BBa_I732095 Colony_2 |
4 | 8/21 BBa_I732095 Colony_3 |
5 | Negative Control |
Liquid Culture
Name | medium |
---|---|
8/20 BBa_I732095 Colony_1 | Plusgrow medium(CP) |
8/20 BBa_I732095 Colony_2 | Plusgrow medium(CP) |
8/20 BBa_I732095 Colony_3 | Plusgrow medium(CP) |
8/22
Miniprep
DNA | Concentration/(µg/ml) | 260/280 | 260/230 |
---|---|---|---|
8/20 BBa_I732095 Colony_1 | 430 | 1.68 | 2.13 |
8/20 BBa_I732095 Colony_2 | 255 | 1.87 | 1.81 |
8/20 BBa_I732095 Colony_3 | 215 | 1.78 | 1.35 |
Restriction Enzyme Digestion
Sample | DNA/(µl) | EcoR1/(µl) | Spe1/(µl) | Buffur/(µl) | MilliQ/(µl) | TOTAL/(µl) |
---|---|---|---|---|---|---|
8/22 BBa_I732095 Colony_1 | 4.7 | 1 | 1 | 3(H) | 20.3 | 30 |
8/22 BBa_I732095 Colony_2 | 7.8 | 1 | 1 | 3(H) | 17.2 | 30 |
8/22 BBa_I732095 Colony_3 | 9.3 | 1 | 1 | 3(H) | 15.7 | 30 |
Transformation
Name | Sample/(µl) | Competent Cells/(µl) | Total/(µl) | Medium |
---|---|---|---|---|
BBa_J04450(pSB3C5) | 2 | 20 | 22 | LB(CP) |
8/25
Transformation
Name | Sample/(µl) | Competent Cells/(µl) | Total/(µl) | Medium |
---|---|---|---|---|
BBa_J04450(pSB2K3) | 2 | 20 | 22 | LB(KAN) |
BBa_C0051(pSB1A2) | 2 | 20 | 22 | LB(AMP) |
Liquid Culture
Name | medium |
---|---|
8/22 BBa_J04450(pSB3C5) | Plusgrow medium(CP)*5 |
8/26
Miniprep
DNA | Concentration/(µg/ml) | 260/280 | 260/230 |
---|---|---|---|
8/25 BBa_J04450(pSB3C5) | 575 | 1.37 | 1.37 |
Liquid Culture
Name | medium |
---|---|
8/25 BBa_J04450(pSB2K3) | Plusgrow medium(KAN)*5 |
8/25 BBa_J0051(pSB1A2) | Plusgrow medium(AMP)*2 |
8/27
Miniprep
DNA | Concentration/(µg/ml) | 260/280 | 260/230 |
---|---|---|---|
8/26 BBa_J04450(pSB2K3) | 1045 | 1.43 | 1.11 |
8/26 BBa_J0051(pSB1A2) | 790 | 1.39 | 0.90 |
Transformation
Name | Sample/(µl) | Competent Cells/(µl) | Total/(µl) | Medium |
---|---|---|---|---|
BBa_J04450(pSB3T5) | 2 | 20 | 22 | LB(TET) |
8/28
Liquid Culture
Name | medium |
---|---|
8/27 BBa_J04450(pSB3T5) | Plusgrow medium(TET) |
8/29
Miniprep
DNA | Concentration/(µg/ml) | 260/280 | 260/230 |
---|---|---|---|
8/28 BBa_J04450(pSB3T5) | 180 | 1.60 | 1.67 |
Transformation
Name | Sample/(µl) | Competent Cells/(µl) | Total/(µl) | Medium |
---|---|---|---|---|
BBa_J04450(pSB4A5) | 2 | 20 | 22 | LB(AMP) |
9/1
Colony PCR
あめふらし
Restriction Enzyme Digestion
Sample | DNA/(µl) | EcoR1/(µl) | Pst1/(µl) | Buffur/(µl) | MilliQ/(µl) | TOTAL/(µl) |
---|---|---|---|---|---|---|
8/28 Miniprep pSB3T5 | 25 | 2 | 2 | 6(H) | 25 | 60 |
Negative control_1 | 3.7 | 0 | 0 | 1(H) | 5.3 | 10 |
8/22 Miniprep I732095 | 14 | 3 | 3 | 9(H) | 61 | 90 |
Negative control_2 | 1.5 | 0 | 0 | 1(H) | 7.5 | 10 |
8/27 Miniprep pSB2K3 | 6 | 3 | 3 | 9(H) | 69 | 90 |
Negative control_3 | 1 | 0 | 0 | 1(H) | 8 | 10 |
8/2 Miniprep Pcon Colony_3 | 20 | 1.5 | 1.5 | 5(H) | 17 | 45 |
Negative control_4 | 4 | 0 | 0 | 1(H) | 5 | 10 |
8/2 Miniprep Pcon Colony_1 | 20 | 1.5 | 1.5 | 5(H) | 17 | 45 |
Negative control_5 | 4 | 0 | 0 | 1(H) | 5 | 10 |
Liquid Culture
Name | medium |
---|---|
8/29 BBa_J04450(pSB4A5) Colony_1 | Plusgrow medium(AMP)*2 |
8/1 Pcon Colony_1 | Plusgrow medium(AMP)*2 |
8/27 pSB3T5 Colony_1 | Plusgrow medium(TET)*2 |
9/2
Miniprep
DNA | Concentration/(µg/ml) | 260/280 | 260/230 |
---|---|---|---|
8/29 BBa_J04450(pSB4A5) Colony_1 | 210 | 1.61 | 1.18 |
8/1 Pcon Colony_1 | 215 | 1.79 | 1.87 |
8/27 pSB3T5 Colony_1 | 215 | 1.58 | 0.92 |
Gel Extractions
Lane | Sample | Volume/(µl) |
---|---|---|
1 | 9/1 pSB3T5 Colony_1 (Nega) | 10 |
2 | 9/1 pSB3T5 Colony_1 (E+P)(Confirmation) | 5 |
3 | 9/1 pSB3T5 Colony_1 (E+P)(Extraction) | 55 |
4 | 1kb ladder | 3 |
5 | 9/1 I732095 (Nega) | 10 |
6 | 9/1 I732095 (E+P)(Confirmation) | 5 |
7 | 9/1 I732095 (E+P)(Extraction) | 85 |
8 | 1kb ladder | 3 |
9 | 9/1 pSB2K3 (Nega) | 10 |
10 | 9/1 pSB2K3 (E+P)(Confirmation) | 5 |
11 | 9/1 pSB2K3 (E+P)(Extraction) | 85 |
Lane | Sample | Volume/(µl) |
---|---|---|
1 | 9/1 Pcon Colony_3 (E+P)(Confirmation) | 5 |
2 | 9/1 Pcon Colony_3 (E+P)(Extraction) | 20 |
3 | 9/1 Pcon Colony_3 (E+P)(Extraction) | 20 |
4 | 9/1 Pcon Colony_3 (Nega) | 10 |
5 | 1kb ladder | 3 |
6 | 9/1 Pcon Colony_1 (E+P)(Confirmation) | 5 |
7 | 9/1 Pcon Colony_1 (E+P)(Extraction) | 20 |
8 | 9/1 Pcon Colony_1 (E+P)(Extraction) | 20 |
9 | 9/1 Pcon Colony_1 (Nega) | 10 |
9/3
Concentration Measurement
DNA | Concentration/(µg/ml) | 260/280 | 260/230 |
---|---|---|---|
pSB2K3 | 40 | 1.48 | 0.10 |
Pcon Colony_1 | 30 | 1.76 | 0.62 |
pSB3T5 | 50 | 2.35 | 0.50 |
pSB1C3 | 65 | 2.14 | 0.10 |
Pcon Colony_3 | 45 | 1.56 | 0.58 |
Ligation
Sample | Insert/(µl) | Vector/(µl) | T4 Ligase/(µl) | 2*Buffer/(µl) | MilliQ/(µl) | Total/(µl) |
---|---|---|---|---|---|---|
Pcon+pSB1C3 | 2.1 | 1.5 | 1 | 5 | 0.4 | 10 |
pSB1C3(Negative Control) | 0 | 1.5 | 1 | 5 | 2.5 | 10 |
Pcon+pSB2K3 | 1.3 | 2.5 | 1 | 5 | 0.2 | 10 |
pSB2K3(Negative Control) | 0 | 2.5 | 1 | 5 | 1.5 | 10 |
Pcon+pSB3T5 | 10 | 2.3 | 1 | 5 | 0.7 | 10 |
pSB3T5(Negative Control) | 0 | 2.3 | 1 | 5 | 1.7 | 10 |
Transformation
Name | Sample/(µl) | Competent Cells/(µl) | SOC Medium/(µl) | Total/(µl) |
---|---|---|---|---|
Pcon+pSB1C3 | 3 | 15 | 72 | 90 |
pSB1C3(Negative Control) | 8 | 10 | 72 | 90 |
Pcon+pSB2K3 | 3 | 15 | 72 | 90 |
pSB2K3(Negative Control) | 8 | 10 | 72 | 90 |
Pcon+pSB3T5 | 3 | 15 | 72 | 90 |
pSB3T5(Negative Control) | 2 | 10 | 48 | 60 |
9/4
Colony PCR
NameⅠ | NameⅡ | NameⅢ | NameⅣ |
---|---|---|---|
- | 9/3 pSB1C3(Nega) Colony_7 | 9/3 Pcon+pSB3T5 Colony_5 | 9/3 Pcon+pSB2K3 Colony_1 |
- | 9/3 pSB1C3(Nega) Colony_8 | 9/3 Pcon+pSB3T5 Colony_6 | 9/3 Pcon+pSB2K3 Colony_2 |
9/3 pSB1C3(Nega) Colony_1 | 9/3 Pcon+pSB3T5 Colony_1 | 9/3 Pcon+pSB3T5 Colony_7 | 9/3 Pcon+pSB2K3 Colony_3 |
9/3 pSB1C3(Nega) Colony_2 | 9/3 Pcon+pSB3T5 Colony_2 | 9/3 Pcon+pSB3T5 Colony_8 | 9/3 Pcon+pSB2K3 Colony_4 |
9/3 pSB1C3(Nega) Colony_3 | 9/3 Pcon+pSB3T5 Colony_3 | 9/3 Pcon+pSB3T5 Colony_9 | 9/3 Pcon+pSB2K3 Colony_5 |
9/3 pSB1C3(Nega) Colony_4 | 9/3 Pcon+pSB3T5 Colony_4 | 9/3 Pcon+pSB3T5 Colony_10 | 9/3 Pcon+pSB2K3 Colony_6 |
9/3 pSB1C3(Nega) Colony_5 | 9/3 pSB1C3(Nega) Colony_9 | 9/3 pSB3T5(Nega) Colony_1 | 9/3 Pcon+pSB2K3 Colony_7 |
9/3 pSB1C3(Nega) Colony_6 | 9/3 pSB1C3(Nega) Colony_10 | 9/3 pSB3T5(Nega) Colony_2 | 9/3 Pcon+pSB2K3 Colony_8 |
NameⅤ | NameⅥ | NameⅦ | NameⅧ |
---|---|---|---|
9/3 Pcon+pSB2K3 Colony_9 | 9/3 Pcon+pSB1C3 Colony_2 | 9/3 Pcon+pSB1C3 Colony_10 | 9/3 pSB1C3(Nega) Colony_8 |
9/3 Pcon+pSB2K3 Colony_10 | 9/3 Pcon+pSB1C3 Colony_3 | 9/3 pSB1C3(Nega) Colony_1 | 9/3 pSB1C3(Nega) Colony_9 |
9/3 pSB2K3(Nega) Colony_1 | 9/3 Pcon+pSB1C3 Colony_4 | 9/3 pSB1C3(Nega) Colony_2 | 9/3 pSB1C3(Nega) Colony_10 |
9/3 pSB2K3(Nega) Colony_2 | 9/3 Pcon+pSB1C3 Colony_5 | 9/3 pSB1C3(Nega) Colony_3 | 9/3 pSB1C3(Nega) Colony_11 |
9/3 pSB2K3(Nega) Colony_3 | 9/3 Pcon+pSB1C3 Colony_6 | 9/3 pSB1C3(Nega) Colony_4 | - |
9/3 pSB2K3(Nega) Colony_4 | 9/3 Pcon+pSB1C3 Colony_7 | 9/3 pSB1C3(Nega) Colony_5 | - |
9/3 pSB2K3(Nega) Colony_5 | 9/3 Pcon+pSB1C3 Colony_8 | 9/3 pSB1C3(Nega) Colony_6 | - |
9/3 Pcon+pSB1C3 Colony_1 | 9/3 Pcon+pSB1C3 Colony_9 | 9/3 pSB1C3(Nega) Colony_7 | - |
NameⅨ | NameⅩ |
---|---|
9/3 Pcon+pSB1C3 Colony_11 | 8/22 Miniprep I732095 Colony_3 |
Negative Control | - |
9/5
Electrophoresis for confirmation
Lane | Restriction Enzyme Digestion Products |
---|---|
1 | Product1 |
2 | Product2 |
3 | Product3 |
4 | Product4 |
5 | Product5 |
6 | 1kbp ladder |