Team:Kyoto/Calendar/24 July 2009

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=='''Lab-note on 24 July 2009'''==
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{| class="notebook"
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==To Do on 24 July==
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|Experimenter||'''Kashima, Takashima, Hibino, Miyazawa'''
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*Miniprep (3-O-2, 3-A-4)
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|-
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*Restriction Enzyme Digestion
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|Note-taker||'''Takashima'''
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*Electrophoresis
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|}
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====Title====
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==Results==
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{| class="notebook"
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===Miniprep===
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|'''24 July: Miniprep Kit, Restriction Enzyme Digestion Electrophoresis'''
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*3-O-2, 3-A-4 was minipreped.
 +
 
 +
===Digestion===
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*Digested with EcoR1 / EcoR1 and Pst1 and run on a gel.
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{| border="1" style="text-align:center;width:50%"
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|+Sample list
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!NO.!!Sample Name!!Enzyme
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|-align="center"
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|1||3-O-2||Eco R1, PST1
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|-align="center"
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|2||3-A-4||Eco R1, PST1
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|-align="center"
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|3||3-O-2||Eco R1
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|-align="center"
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|4||3-A-4||Eco R1
|}
|}
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====Main Protocol====
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===Electrophoresis===
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{| class="notebook"
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[[Image:Kyoto_e_090724_1.png|250px|right]]
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|
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* Very clear bands were found. (See the gel image in right!)
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#[[#Miniprep|Miniprep Kit]]
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* The bands are listed below.
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#Restriction Enzyme Digestion
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#[[#Electrophoresis|Electrophoresis]]
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{| border="1" style="text-align:center;width:50%"
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|}
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|+Parts and Plasmid length
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====Storage list====
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!!!3-O-2!!3-A-4
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{| class="notebook"
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!Sample Date!!Sample Name!!Enzyme!!purification Kit!!Product!!Storage
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|-
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|iGEM DNA parts kit||3-O-2||Eco R1|| || ||6F freezer
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|-
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|iGEM DNA parts kit||3-A-4||Eco R1|| || ||6F freezer
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|-
|-
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|iGEM DNA parts kit||3-O-2||Eco R1<br>PST1|| || ||6F freezer
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|Parts length / bp||2837||487
|-
|-
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|iGEM DNA parts kit||3-A-4||Eco R1<br>PST1|| || ||6F freezer
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|Plasmid length / bp||3266||3266
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|}
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====Miniprep====
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{| class="notebook"
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|
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'''Protocol'''
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*[3-O-2, 3-A-4]
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#Cultured samples (090722 9:30 - 090723 16:50)
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#Centrifuged 2 mins at 15000 G 4 °C.
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#Removed the top of layer.
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#Centrifuged 2 mins at 15000 G 4 °C.
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#Removed the top of layer.
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#Centrifuged 10 mins at 14000 G 4 °C.
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#Applied  the top of layer to column.
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#Centrifuged RT, 10 sec and remove through.
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#Added Buffer PB 500 µl.
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#Centrifuged RT, 10 sec and remove through.
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#Added Buffer PE 650 µl.
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#Centrifuged RT, 10 sec and remove through.
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#Centrifuged 1min at 17900 G 4 °C.
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#Exchanged tube. (remove Ethanol)
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#Added Milli Q 50 µl RT 1min.
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#Centrifuged 1min at 17900 G 4°C.
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#Collected through and measured concentration.
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----
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'''Results'''
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{| class="data"
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!Sample!!conc./(ng/µl)
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|-
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|3-O-2||283.5
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|-
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|3-A-4||103.9
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|}
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|}
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====Electrophoresis====
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{| class="notebook"
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|
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'''Protocol'''
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*Gel:'''Agarose'''
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*Time:'''12:50-13:20'''
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*Voltage:'''100 V'''
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{| class="data"
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! !!1!!2!!3!!4
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|-
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!Sample
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|3-O-2 (3 µl)||3-A-4 (3 µl)||3-O-2 (3 µl)||3-A-4 (3 µl)
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|-
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!enzyme
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|Eco R1 (0.17 µl)||Eco R1 (0.17 µl)||Eco R1 (0.17 µl)||Eco R1 (0.17 µl)
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|-
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!enzyme
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|PST1 (0.3 µl)||PST1 (0.3 µl)|| ||
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|-
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!Water / µl
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|1 ||1 ||1.4||1.4
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|-
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!H buffer / µl
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|8.2||8.2||8.1||8.1
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|-
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!Total / µl
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!12.7!!12.7!!12.9!!12.9
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|}
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Incubated at 37 °C (090724 12:50 to 13:20 )
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{| class="data"
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! !!1!!2!!3!!4!!5
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|-
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!Sample
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|3-O-2||3-A-4||3-O-2||3-A-4||Marker
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|-
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!volume / µl
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|10||10||13||13||2.5
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|-
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!6 times loddingbuffer / µl
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|2||2||2.6||2.6||0.5
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|}
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----
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'''Results'''
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[[Image:Kyoto_e_090724_1.png|thumb|300px|Result of Electrophoresis.]]
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{| class="data"
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! !!3-O-2!!3-A-4
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|-
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!Parts length / bp
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|2837||487
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|-
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!Plasmid length / bp
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|3266||3266
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|}
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----
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'''Discussion'''
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*This result indicates that the parts ( 3-O-2, 3-A-4) were  treated with  enzyme in appropriate way.
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|}
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Latest revision as of 06:53, 14 August 2009

March
MTWTFSS
            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
April
MTWTFSS
    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
May
MTWTFSS
        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
June
MTWTFSS
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
July
MTWTFSS
    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
MTWTFSS
          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
MTWTFSS
  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
October
MTWTFSS
      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

To Do on 24 July

  • Miniprep (3-O-2, 3-A-4)
  • Restriction Enzyme Digestion
  • Electrophoresis

Results

Miniprep

  • 3-O-2, 3-A-4 was minipreped.

Digestion

  • Digested with EcoR1 / EcoR1 and Pst1 and run on a gel.
Sample list
NO.Sample NameEnzyme
13-O-2Eco R1, PST1
23-A-4Eco R1, PST1
33-O-2Eco R1
43-A-4Eco R1

Electrophoresis

Kyoto e 090724 1.png
  • Very clear bands were found. (See the gel image in right!)
  • The bands are listed below.
Parts and Plasmid length
3-O-23-A-4
Parts length / bp2837487
Plasmid length / bp32663266

Retrieved from "http://2009.igem.org/Team:Kyoto/Calendar/24_July_2009"