Team:Chiba/jk/β/week 1

From 2008.igem.org

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>Protocols


Contents

Week 1

17 August, 2008

Transformation
competent cells : XL10G
  • BBa_R0040(2008) [http://partsregistry.org/Part:BBa_R0040] -> No colonies on plates
  • BBa_C0070(2008) [http://partsregistry.org/Part:BBa_C0070] -> No colonies on plates
  • BBa_C0076(2008) [http://partsregistry.org/Part:BBa_C0076] -> No colonies on plates
  • BBa_C0077(2008) [http://partsregistry.org/Part:BBa_C0077] -> No colonies on plates
  • BBa_C0078(2008) [http://partsregistry.org/Part:BBa_C0078] -> No colonies on plates
  • BBa_C0079(2008) [http://partsregistry.org/Part:BBa_C0079] -> No colonies on plates

18 August, 2008

PCR
  • BBa_C0070(2008) [http://partsregistry.org/Part:BBa_C0070]
  • BBa_C0076(2008) [http://partsregistry.org/Part:BBa_C0076]
  • BBa_C0077(2008) [http://partsregistry.org/Part:BBa_C0077]
  • BBa_C0078(2008) [http://partsregistry.org/Part:BBa_C0078]
  • BBa_C0079(2008) [http://partsregistry.org/Part:BBa_C0079]
  • BBa_F2620(2007) [http://partsregistry.org/Part:BBa_F2620]・・・control


DNA Template 1
dNTP mix 10
Foward Primer 5
Reverse Primer 5
DNA polymerase 1
Thermopol Buffer 10
dH2O 68
TOTAL 100μL


95℃,5min -> ( 95℃,1min -> 52℃,1min -> 72℃,1min )・・・25cycles -> 72℃,10min -> 6℃


--->Gel Check


Chiba-0818.JPG
Sample DNA 3
Loading Dye 2
dH2O 7
TOTAL 12μL
From left;
  • BBa_C0070(2008) [http://partsregistry.org/Part:BBa_C0070] ->None
  • BBa_C0076(2008) [http://partsregistry.org/Part:BBa_C0076] ->None
  • BBa_C0077(2008) [http://partsregistry.org/Part:BBa_C0077] ->None
  • BBa_C0078(2008) [http://partsregistry.org/Part:BBa_C0078] ->None
  • BBa_C0079(2008) [http://partsregistry.org/Part:BBa_C0079] ->None
  • BBa_F2620(2007) [http://partsregistry.org/Part:BBa_F2620]・・・control ->OK



Transformation

competent cells : XL10G

  • BBa_C0070(2007) [http://partsregistry.org/Part:BBa_C0070]
  • BBa_C0070(2006) [http://partsregistry.org/Part:BBa_C0070]
  • BBa_C0076(2007) [http://partsregistry.org/Part:BBa_C0076]
  • BBa_C0076(2006) [http://partsregistry.org/Part:BBa_C0076] -> No colonies on plates(19/8)
  • BBa_C0078(2007) [http://partsregistry.org/Part:BBa_C0078]
  • BBa_C0078(2006) [http://partsregistry.org/Part:BBa_C0078]
  • BBa_C0170(2007) [http://partsregistry.org/Part:BBa_C0170]
  • BBa_C0170(2006) [http://partsregistry.org/Part:BBa_C0170]
  • BBa_C0178(2007) [http://partsregistry.org/Part:BBa_C0178]
  • BBa_C0178(2006) [http://partsregistry.org/Part:BBa_C0178]


--->(20/8)Mini prep

19 August, 2008

PCR
  • BBa_I9026(2007) [http://partsregistry.org/Part:BBa_I9026]
  • BBa_I9026(2006) [http://partsregistry.org/Part:BBa_I9026]
  • BBa_I9030(2007) [http://partsregistry.org/Part:BBa_I9030]
  • BBa_I9030(2006) [http://partsregistry.org/Part:BBa_I9030]
  • BBa_S3154(2007) [http://partsregistry.org/Part:BBa_S3154]
  • BBa_S3154(2006) [http://partsregistry.org/Part:BBa_S3154]
  • BBa_F2620(2007) [http://partsregistry.org/Part:BBa_F2620]・・・control


DNA Template 1
dNTP mix 10
Foward Primer 5
Reverse Primer 5
DNA polymerase VENT 1
Thermopol Buffer 5
dH2O 28
TOTAL 50μL


95℃,5min -> ( 95℃,1min -> 52℃,1min -> 72℃,1min )・・・25cycles -> 72℃,10min -> 6℃


--->Gel Check
Chiba-0819.JPG
Sample DNA 3
Loading Dye 2
dH2O 7
TOTAL 12μL
From left;
  • BBa_I9026(2007) [http://partsregistry.org/Part:BBa_I9026] ->None
  • BBa_I9026(2006) [http://partsregistry.org/Part:BBa_I9026] ->None
  • BBa_I9030(2007) [http://partsregistry.org/Part:BBa_I9030] ->None
  • BBa_I9030(2006) [http://partsregistry.org/Part:BBa_I9030] ->None
  • BBa_S3154(2007) [http://partsregistry.org/Part:BBa_S3154] ->None
  • BBa_S3154(2006) [http://partsregistry.org/Part:BBa_S3154] ->None
  • BBa_F2620(2007) [http://partsregistry.org/Part:BBa_F2620]・・・control ->OK


20 August, 2008

(18/8)--->Mini prep
  • BBa_C0070(2007) [http://partsregistry.org/Part:BBa_C0070]
  • BBa_C0070(2006) [http://partsregistry.org/Part:BBa_C0070]
  • BBa_C0076(2007) [http://partsregistry.org/Part:BBa_C0076]
  • BBa_C0078(2007) [http://partsregistry.org/Part:BBa_C0078]
  • BBa_C0078(2006) [http://partsregistry.org/Part:BBa_C0078]
  • BBa_C0170(2007) [http://partsregistry.org/Part:BBa_C0170]
  • BBa_C0170(2006) [http://partsregistry.org/Part:BBa_C0170]
  • BBa_C0178(2007) [http://partsregistry.org/Part:BBa_C0178]
  • BBa_C0178(2006) [http://partsregistry.org/Part:BBa_C0178]


--->Gel Check
Chiba-0820-3.JPG
Sample DNA 3
Loading Dye 2
dH2O 7
TOTAL 12μL
From left;
  • BBa_C0076(2007) [http://partsregistry.org/Part:BBa_C0076] -> None
  • BBa_C0078(2007) [http://partsregistry.org/Part:BBa_C0078] -> None
  • BBa_C0078(2006) [http://partsregistry.org/Part:BBa_C0078] -> None


Chiba-0820-4.JPG
Sample DNA 10
Loading Dye 4
dH2O 10
TOTAL 24μL
From left;
  • BBa_C0078(2007) [http://partsregistry.org/Part:BBa_C0078] -> None
  • BBa_C0078(2006) [http://partsregistry.org/Part:BBa_C0078] -> OK
  • BBa_C0076(2007) [http://partsregistry.org/Part:BBa_C0076] -> None
  • BBa_C0070(2007) [http://partsregistry.org/Part:BBa_C0070] -> OK
  • BBa_C0070(2006) [http://partsregistry.org/Part:BBa_C0070] -> OK



Chiba-0820.JPG
Sample DNA 3
Loading Dye 2
dH2O 7
TOTAL 12μL
From left;
  • BBa_C0170(2007) [http://partsregistry.org/Part:BBa_C0170] -> OK
  • BBa_C0170(2006) [http://partsregistry.org/Part:BBa_C0170] -> OK
  • BBa_C0178(2007) [http://partsregistry.org/Part:BBa_C0178] -> OK
  • BBa_C0178(2006) [http://partsregistry.org/Part:BBa_C0178] -> OK


--->Digestion test

  • BBa_C0170(2007) [http://partsregistry.org/Part:BBa_C0170]
  • BBa_C0170(2006) [http://partsregistry.org/Part:BBa_C0170]
  • BBa_C0178(2007) [http://partsregistry.org/Part:BBa_C0178]
  • BBa_C0178(2006) [http://partsregistry.org/Part:BBa_C0178]
Sample No 12345678
BBa_C0170(2006) [http://partsregistry.org/Part:BBa_C0170] 3---3---
BBa_C0170(2007) [http://partsregistry.org/Part:BBa_C0170] -3---3--
BBa_C0178(2006) [http://partsregistry.org/Part:BBa_C0178] --3---3-
BBa_C0178(2007) [http://partsregistry.org/Part:BBa_C0178] ---3---3
EcoRⅠ 0.10.10.10.10.10.10.10.1
PstⅠ ----0.10.10.10.1
Buffer EcoRⅠ 11111111
BSA 11111111
dH2O 4.94.94.94.94.84.84.84.8
TOTAL 1010101010101010


--->Gel Check
Chiba-0820-2.JPG
Sample DNA 3
Loading Dye 2
dH2O 7
TOTAL 12μL
From left;
(Single Digestion 1~4)
  • BBa_C0170(2006) [http://partsregistry.org/Part:BBa_C0170] -> OK
  • BBa_C0170(2007) [http://partsregistry.org/Part:BBa_C0170] -> OK
  • BBa_C0178(2006) [http://partsregistry.org/Part:BBa_C0178] -> OK
  • BBa_C0178(2007) [http://partsregistry.org/Part:BBa_C0178] -> OK
(Double Digestion 1~4)
  • BBa_C0170(2006) [http://partsregistry.org/Part:BBa_C0170] -> OK
  • BBa_C0170(2007) [http://partsregistry.org/Part:BBa_C0170] -> OK
  • BBa_C0178(2006) [http://partsregistry.org/Part:BBa_C0178] -> OK
  • BBa_C0178(2007) [http://partsregistry.org/Part:BBa_C0178] -> OK

21 August, 2008

(20/8)--->PCR
  • BBa_C0070(2007) [http://partsregistry.org/Part:BBa_C0070]
  • BBa_C0070(2006) [http://partsregistry.org/Part:BBa_C0070]
  • BBa_C0076(2007) [http://partsregistry.org/Part:BBa_C0076]
  • BBa_C0078(2007) [http://partsregistry.org/Part:BBa_C0078]
  • BBa_C0078(2006) [http://partsregistry.org/Part:BBa_C0078]


DNA Template 1
dNTP mix 10
Foward Primer 5
Reverse Primer 5
DNA polymerase TAQ 1
Thermopol Buffer 5
dH2O 28
TOTAL 50μL


95℃,5min -> ( 95℃,1min -> 52℃,1min -> 72℃,1min )・・・25cycles -> 72℃,10min -> 6℃


--->Gel Check
Chiba-0821.JPG
Sample DNA 3
Loading Dye 2
dH2O 7
TOTAL 12μL
From left;
  • BBa_C0078(2007) [http://partsregistry.org/Part:BBa_C0078] -> None
  • BBa_C0078(2006) [http://partsregistry.org/Part:BBa_C0078] -> None
  • BBa_C0076(2007) [http://partsregistry.org/Part:BBa_C0076] -> None
  • BBa_C0070(2007) [http://partsregistry.org/Part:BBa_C0070] -> OK
  • BBa_C0070(2006) [http://partsregistry.org/Part:BBa_C0070] -> OK


Transformation
competent cells : XL10G
  • BBa_S03154(2007) [http://partsregistry.org/Part:BBa_S03154]
  • BBa_S03154(2006) [http://partsregistry.org/Part:BBa_S03154]
  • BBa_I9026(2007) [http://partsregistry.org/Part:BBa_I9026]
  • BBa_I9026(2006) [http://partsregistry.org/Part:BBa_I9026]
  • BBa_I9030(2007) [http://partsregistry.org/Part:BBa_I9030]
  • BBa_I9030(2006) [http://partsregistry.org/Part:BBa_I9030]



--->(23/8)Digestion


22 August, 2008

Transformation
competent cells : XL10G
  • BBa_J13002(2007) [http://partsregistry.org/Part:BBa_J13002]
  • BBa_J13002(2006) [http://partsregistry.org/Part:BBa_J13002]
  • BBa_J04500(2007) [http://partsregistry.org/Part:BBa_J04500]
  • BBa_J04500(2006) [http://partsregistry.org/Part:BBa_J04500]
(Locate & 2 Punch ~iyama ver.~)
  • BBa_C0076(2008) [http://partsregistry.org/Part:BBa_C0076] -> No colonies on plates
  • BBa_C0077(2008) [http://partsregistry.org/Part:BBa_C0077] -> No colonies on plates
  • BBa_C0078(2008) [http://partsregistry.org/Part:BBa_C0078] -> No colonies on plates
  • BBa_J04500(2008) [http://partsregistry.org/Part:BBa_J04500] -> No colonies on plates


--->(24/8)Mini prep

23 August, 2008

(21/8)--->Mini prep
  • BBa_S03154(2007) [http://partsregistry.org/Part:BBa_S03154]
  • BBa_S03154(2006) [http://partsregistry.org/Part:BBa_S03154]
  • BBa_I9026(2007) [http://partsregistry.org/Part:BBa_I9026]
  • BBa_I9026(2006) [http://partsregistry.org/Part:BBa_I9026]
  • BBa_I9030(2007) [http://partsregistry.org/Part:BBa_I9030]
  • BBa_I9030(2006) [http://partsregistry.org/Part:BBa_I9030]


--->Digestion test
  • BBa_S03154(2007) [http://partsregistry.org/Part:BBa_S03154]
  • BBa_S03154(2006) [http://partsregistry.org/Part:BBa_S03154]
  • BBa_I9026(2007) [http://partsregistry.org/Part:BBa_I9026]
  • BBa_I9026(2006) [http://partsregistry.org/Part:BBa_I9026]
  • BBa_I9030(2007) [http://partsregistry.org/Part:BBa_I9030]
  • BBa_I9030(2006) [http://partsregistry.org/Part:BBa_I9030]
  • BBa_J04500(2007) [http://partsregistry.org/Part:BBa_J04500]


Sample No 1 2 3 4 5 6 7 8 9101112131415161718192021
BBa_I9030(2007) [http://partsregistry.org/Part:BBa_I9030] 3------3------3------
BBa_I9030(2006) [http://partsregistry.org/Part:BBa_I9030] -3------3------3-----
BBa_S03154(2007) [http://partsregistry.org/Part:BBa_S03154] --3------3------3----
BBa_S03154(2006) [http://partsregistry.org/Part:BBa_S03154] ---3------3------3---
BBa_I9026(2007) [http://partsregistry.org/Part:BBa_I9026] ----3------3------3--
BBa_I9026(2007) [http://partsregistry.org/Part:BBa_I9026] -----3------3------3-
BBa_J04500(2007) [http://partsregistry.org/Part:BBa_J04500] ------1------3------3
EcoRⅠ -------0.10.10.10.10.10.10.10.10.10.10.10.10.10.1
PstⅠ --------------0.10.10.10.10.10.10.1
Buffer EcoRⅠ -------0.90.90.90.90.90.90.90.80.80.80.80.80.80.8
BSA -------11111111111111
dH2O -------55555555555555
TOTAL 33333311010101010101010101010101010


--->Gel Check
Chiba-0823-1.JPG
Sample No 1~67
Sample DNA 31
Loading Dye 11
dH2O 24
TOTAL 66
From left; Sanple No. 1~7
  • BBa_S03154(2007) [http://partsregistry.org/Part:BBa_S03154]
  • BBa_S03154(2006) [http://partsregistry.org/Part:BBa_S03154]
  • BBa_I9026(2007) [http://partsregistry.org/Part:BBa_I9026]
  • BBa_I9026(2006) [http://partsregistry.org/Part:BBa_I9026]
  • BBa_I9030(2007) [http://partsregistry.org/Part:BBa_I9030]
  • BBa_I9030(2006) [http://partsregistry.org/Part:BBa_I9030]
  • BBa_J04500(2007) [http://partsregistry.org/Part:BBa_J04500]



Chiba-0823-2.JPG
Sample No 8~21
Sample DNA 10
Loading Dye 2
TOTAL 12
From left;
(Single Digestion) Sanple No. 8~14
  • BBa_S03154(2007) [http://partsregistry.org/Part:BBa_S03154] -> OK
  • BBa_S03154(2006) [http://partsregistry.org/Part:BBa_S03154] -> OK
  • BBa_I9026(2007) [http://partsregistry.org/Part:BBa_I9026] -> OK
  • BBa_I9026(2006) [http://partsregistry.org/Part:BBa_I9026] -> OK
  • BBa_I9030(2007) [http://partsregistry.org/Part:BBa_I9030] -> OK
  • BBa_I9030(2006) [http://partsregistry.org/Part:BBa_I9030] -> OK
  • BBa_J04500(2007) [http://partsregistry.org/Part:BBa_J04500] -> OK
(Double Digestion) Sample No. 15~21
  • BBa_S03154(2007) [http://partsregistry.org/Part:BBa_S03154] -> OK
  • BBa_S03154(2006) [http://partsregistry.org/Part:BBa_S03154] -> OK
  • BBa_I9026(2007) [http://partsregistry.org/Part:BBa_I9026] -> OK
  • BBa_I9026(2006) [http://partsregistry.org/Part:BBa_I9026] -> OK
  • BBa_I9030(2007) [http://partsregistry.org/Part:BBa_I9030] -> OK
  • BBa_I9030(2006) [http://partsregistry.org/Part:BBa_I9030] -> OK
  • BBa_J04500(2007) [http://partsregistry.org/Part:BBa_J04500] -> OK?



Digestion test
BBa_J04500(2007:old-sample)[http://partsregistry.org/Part:BBa_J04500]
Sample No 123
Sample DNA 333
EcoRⅠ -0.10.1
SpeⅠ --0.1
EcoRⅠ Buffer -0.90.8
BSA -11
dH2O -55
TOTAL 31010


--->Gel Check

Chiba-0823-3.JPG
Sample No 123
Sample DNA 31010
Loading Dye 122
dH2O 2--
TOTAL 61212
From left;
  1. BBa_J04500(2007) [http://partsregistry.org/Part:BBa_J04500] -> OK
  2. BBa_J04500(2007) [http://partsregistry.org/Part:BBa_J04500]Single Digestion -> OK
  3. BBa_J04500(2007) [http://partsregistry.org/Part:BBa_J04500]Double Digestion -> Bad

>next week

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