Team:Kyoto/Notebook

From 2014.igem.org

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</table>
</table>
 +
<h2>8/29</h2>
 +
<h3>Miniprep</h3>
 +
<table class="note">
 +
<tr><th>DNA</th><th>Concentration(&micro;g/ml)</th><th>260/280</th><th>260/230</th></tr>
 +
<tr><td>7/16 BBa_B0030-(1)</td><td>229</td><td>1.96</td><td>1.40</td></tr>
 +
</table>
 +
<h3>transformation</h3>
 +
<table class="note">
 +
<tr><th>Name</th><th>Sample(&micro;l)</th><th>Competent Cells(&micro;l)</th><th>Total(&micro;l)</th><th>Plate</th></tr>
 +
<tr><td>BBa_J04450</td><td>2</td><td>20(2/23)</td><td>22</td><td>LB（CP）</td></tr>
 +
</table>
 +
 +
<h2>9/1</h2>
 +
<h3>Colony PCR</h3>
 +
 +
<h3>Restriction Enzyme Digestion</h3>
 +
<table class="note">
 +
<tr><th>7/18 RBS Sample</th><th>EcoR1</th><th>Xba1</th><th>Spe1</th><th>Pst1</th><th>Buffur</th><th>BSA</th><th>MilliQ</th><th>TOTAL</tr>
 +
<tr><td>Sample1</td><td>0.3</td><td>0</td><td>0</td><td>0</td><td>1(H)</td><td>0</td><td>7.7</td><td>10</td></tr>
 +
<tr><td>Control1</td><td>0</td><td>0</td><td>0</td><td>0</td><td>1</td><td>0</td><td>8</td><td>10</td></tr>
 +
<tr><td>Sample2</td><td>0</td><td>0.3</td><td>0</td><td>0</td><td>1(M)</td><td>1</td><td>6.7</td><td>10</td></tr>
 +
<tr><td>Control2</td><td>0</td><td>0</td><td>0</td><td>0</td><td>1</td><td>0</td><td>8</td><td>10</td></tr>
 +
<tr><td>Sample3</td><td>0</td><td>0</td><td>0.3</td><td>0</td><td>1(M)</td><td>0</td><td>7.7</td><td>10</td></tr>
 +
<tr><td>Control3</td><td>0</td><td>0</td><td>0</td><td>0</td><td>1</td><td>0</td><td>8</td><td>10</td></tr>
 +
<tr><td>Sample4</td><td>0</td><td>0</td><td>0</td><td>0.3</td><td>1(H)</td><td>0</td><td>7.7</td><td>10</td></tr>
 +
<tr><td>Control4</td><td>0</td><td>0</td><td>0</td><td>0</td><td>1</td><td>0</td><td>8</td><td>10</td></tr>
 +
<tr><td>Control5</td><td>0</td><td>0</td><td>0</td><td>0</td><td>1</td><td>0</td><td>8</td><td>10</td></tr>
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</table>
</div>
</div>
</html>
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Revision as of 01:27, 2 September 2014

  1. REDIRECT Template:Kyoto/css
count down

Home Team Official Team Profile Project Parts Modeling Notebook Safety Attributions

Notebook

You should make use of the calendar feature on the wiki and start a lab notebook. This may be looked at by the judges to see how your work progressed throughout the summer. It is a very ustoeful organizational tool as well.

July
Sun.Mon.Tues.Wed.Thurs.Fri.Sat.


12345
6789101112
13141516171819
20212223242526
2728293031
August
Sun.Mon.Tues.Wed.Thurs.Fri.Sat.





12
3456789
10111213141516
17181920212223
24252627282930
31
September
Sun.Mon.Tues.Wed.Thurs.Fri.Sat.

123456
78910111213
14151617181920
21222324252627
282930

7/15

transformation

NameSample(µl)Competent Cells(µl)Total(µl)Plate
BBa_J23101220(2/23)22LB（CP）
BBA_B0032220（2/23）22LB（CP）

7/16

transformation

NameSample(µl)Competent Cells(µl)Total(µl)Plate
BBa_B0030220(2/23)22LB（CP）
BBA_B0032220（2/23）22LB（CP）

7/17

Colony PCR

Sample
7/16 BBa_B0030-(1)
7/16 BBa_B0030-(2)
7/16 BBa_B0030-(3)
7/16 BBa_B0030-(4)
7/16 BBa_B0030-(5)
7/16 BBa_B0030-(6)
7/16 BBa_B0032-(1)
7/16 BBa_B0032-(2)
7/16 BBa_B0032-(3)
7/16 BBa_B0032-(4)
7/16 BBa_B0032-(5)
7/16 BBa_B0032-(6)
NC

Liquid Culture

Samplemedium
7/16 BBa_B0030-(1)Plusgrow medium(+CP)
7/16 BBa_B0030-(2)Plusgrow medium(+CP)
7/16 BBa_B0030-(3)Plusgrow medium(+CP)
7/16 BBa_B0030-(4)Plusgrow medium(+CP)
7/16 BBa_B0030-(5)Plusgrow medium(+CP)
7/16 BBa_B0030-(6)Plusgrow medium(+CP)
7/16 BBa_B0032-(1)Plusgrow medium(+CP)
7/16 BBa_B0032-(2)Plusgrow medium(+CP)
7/16 BBa_B0032-(3)Plusgrow medium(+CP)
7/16 BBa_B0032-(4)Plusgrow medium(+CP)
7/16 BBa_B0032-(5)Plusgrow medium(+CP)
7/16 BBa_B0032-(6)Plusgrow medium(+CP)

7/18

Miniprep

DNAConcentration(µg/ml)260/280260/230
7/16 BBa_B0030-(1)2291.961.40
7/16 BBa_B0030-(2)2091.951.50
7/16 BBa_B0030-(3)1082.122.12
7/16 BBa_B0030-(4)186--
7/16 BBa_B0030-(5)1742.021.56
7/16 BBa_B0030-(6)1582.011.68
7/16 BBa_B0032-(1)1792.002.06
7/16 BBa_B0032-(3)2211.901.37
7/16 BBa_B0032-(4)2251.891.37
7/16 BBa_B0032-(5)2701.981.63
7/16 BBa_B0032-(6)1941.891.33

7/28

Restriction Enzyme Digestion

7/18 RBS SampleEcoR1Xba1Spe1Pst1BuffurBSAMilliQTOTAL
Sample10.30001(H)07.710
Sample200.3001(M)16.710
Sample3000.301(M)07.710
Sample40000.31(H)07.710
Control000010810

7/30

Restriction Enzyme Digestion

7/18 RBS SampleEcoR1Xba1Spe1Pst1BuffurBSAMilliQTOTAL
Sample10.30001(H)07.710
Sample200.3001(M)16.710
Sample3000.301(M)07.710
Sample40000.31(H)07.710
Control000010810

7/30

Electrophoresis for confirmation

LaneRestriction Enzyme Digestion Products
1Sample1
2Sample2
3Sample3
4Sample4
5Sample5
61kbp ladder

7/31

Transformation

2-24B BBa_E0240(RBS-GFP-DT)+CP4-17D J23100(Pcon) +Amp
plasmid2μL2μL
DH5α20μL2μL
TOTAL22μL22μL

7/31

Restriction Enzyme Digestion

SamplePlasmidEcoR1Xba1Spe1Pst1BuffurBSAMilliQTOTAL
Sample150.30001(H)03.710
Sample2500.3001(M)03.710
Sample35000.301(H)03.710
Sample450000.31(M)12.710
Control500001(M)0410

8/1

Electrophoresis for confirmation

LaneSample
11kb ladder
27/31 Restruction Enzyme Digestion Products Sample1
37/31 Restruction Enzyme Digestion Products Sample2 Error
4NC
5NC
67/31 Restruction Enzyme Digestion Products Sample3
77/31 Restruction Enzyme Digestion Products Sample4
87/31 Restruction Enzyme Digestion Products Sample5

8/1

PCR

BBa_E0240(RBS-GFP-DT)
Volume
10×buffur5μL
2mMdNTPs5μL
25mM MgSO42μL
VF Primer1.5μL
VR Primer1.5μL
template0.5μL
KOD-plus1μL
MilliQ33.5μL
TOTAL50μL

8/1

PCR

BBa_E0240(RBS-GFP-DT)
TemparatureTime
Pre Denature94℃2min
Denature94℃15sec30cycle
Anniling55℃30sec30cycle
Exensim68℃1min30cycle

8/1

densitomitory after column prification

BBa_E0240(RBS-GFP-DT)
Density260/230260/280
80μg/mL1.781.91

8/2

Miniprep

7/31 Pcon
SampleDensity(μg/mL)260/280260/230
12.92.113.09
21.32.59-
33.22.013.13
42.81.872.70
52.12.395.90
63.61.901.37

8/20

Transformation

BBa_I7320957/31 Transformation BBa_E0240(RBS-aFD-DT)(+CP)
plasmid2μL2μL
DH5α20μL20μL
TOTAL22μL22μL
×2×2

8/21

Colony PCR

Master Mix
Quick Taq20μL
VF0.8μL
VR0.8μL
MilliQ18.4μL
TOTAL40μL

8/21

Colony PCR

TemparatureTime
Pre Denature94.0℃2min
Denature94.0℃30sec
Anniling55.0℃0.8μL
Extention68.0℃18.4μL
Denature - Extention 30 cycle

8/21

Electrophoresis for confirmation

LaneSample
11kb ladder
2BBa_E0430-Sample1
3BBa_E0430-Sample2
4BBa_E0430-Sample3
5Negative Control

8/21

Electrophoresis for confirmation

1BBa_E0430-Sample1
2BBa_E0430-Sample2
3BBa_E0430-Sample3

8/22

Miniprep

Sampledensity260/280260/230
BBa_I732095-Sample1430µg/mL1.682.13
BBa_I732095-Sample2255µg/mL1.871.81
BBa_I732095-Sample3215µg/mL1.781.35

8/22

Restriction Enzyme Digestion

BBa_I732095-Sample14.7µL
EcoRI1µL
SpeRI1µL
Buffur(H)3µL
MilliQ20.3µL
TOTAL30µL
BBa_I732095-Sample27.8µL
EcoRI1µL
SpeRI1µL
Buffur(H)3µL
MilliQ20.3µL
TOTAL30µL
BBa_I732095-Sample39.3µL
EcoRI1µL
SpeRI1µL
Buffur(H)3µL
MilliQ15.7µL
TOTAL30µL

8/22

Transformation

BBa_J04450(pSB3C5)
DNA2µL
Conpitent cell20µL

8/22

Transformation

BBa_J04450(pSB3C5)Time
on ice30min
heat shock45sec
on ice2min
Preculture1hour

8/25

Transformation

BBa_J04450(2014)
DNA20µL
Conpitent cell40µL
BBa_J04450(2013)
DNA20µL
Conpitent cell40µL
BBa_C0051
DNA20µL
Conpitent cell40µL

8/25

liquid culture

8/22 J04450(psB3C5) 30mL

8/26

Miniprep

Sampledensity(µg/mL)260/280260/230
J04450(psB3C5)5751.371.34

8/26

liquid culture

8/25 psB2K3×5(30mL)
8/25 psB1A2×2(12mL)

8/27

Miniprep

Sampledensity260/280260/230
8/26 licuid culture psB2K31045(µg/mL)1.431.11
8/26 licuid culture psB1A2790(µg/mL)1.390.90

8/27

BBa_J04450(psB3T5)

DNA6mL
Tet60µg(10µg/mL)

8/29

Miniprep

DNAConcentration(µg/ml)260/280260/230
7/16 BBa_B0030-(1)2291.961.40

transformation

NameSample(µl)Competent Cells(µl)Total(µl)Plate
BBa_J04450220(2/23)22LB（CP）

9/1

Colony PCR

Restriction Enzyme Digestion

7/18 RBS SampleEcoR1Xba1Spe1Pst1BuffurBSAMilliQTOTAL
Sample10.30001(H)07.710
Control1000010810
Sample200.3001(M)16.710
Control2000010810
Sample3000.301(M)07.710
Control3000010810
Sample40000.31(H)07.710
Control4000010810
Control5000010810

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