Team:Chiba/Notebook/Calendar/17 September 2009

From 2009.igem.org

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Single Digestion
Single Digestion
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{| border="1" align="center"
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{| border="1"  
| || volume (uL)
| || volume (uL)
|- align="center"
|- align="center"
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{| border="1" align=
{| border="1" align=
|- align="center"
|- align="center"
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| sample number || 1 || 2 || 3 || 4 || 5 || 6 || 7 || 8 || 9 || 10 || 11 || 12 || 13 || 14 ||15 || 16  
+
| sample number || (1) || (2) || (3) || (4) || (5) || (6) || (7) || (8) || (9) || (10) || (11) || (12) || (13) || (14) || (15) || (16)
|- align="center"
|- align="center"
|
|
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| 5x sequencing Buffer || 1|| 1 || 1 || 1 || 1 || 1 || 1 || 1 || 1 || 1 || 1 || 1 || 1 || 1 || 1 || 1
| 5x sequencing Buffer || 1|| 1 || 1 || 1 || 1 || 1 || 1 || 1 || 1 || 1 || 1 || 1 || 1 || 1 || 1 || 1
|- align="center"
|- align="center"
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| PstI || 0.20 (4U)
+
| LuxR 3 (Mutant number) || 1 || 1 ||
|- align="center"
|- align="center"
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| Buffer(10x) || 1
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| 4 |||||| 1 || 1
|- align="center"
|- align="center"
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| BSA(10x)|| 1
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| 6 |||||||||| 1 || 1 ||
|- align="center"
|- align="center"
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| dW || 4.6
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| 7 |||||||||||||| 1 || 1
|- align="center"
|- align="center"
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| total || 10
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| 9 |||||||||||||||||| 1 || 1
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|- align="center"
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| 10 |||||||||||||||||||||| 1 || 1
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|- align="center"
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| 11 |||||||||||||||||||||||||| 1 || 1
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|- align="center"
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| L1 |||||||||||||||||||||||||||||| 1 || 1
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|- align="center"
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| Primer(VF) || 0.5 |||| 0.5 |||| 0.5 |||| 0.5 |||| 0.5 |||| 0.5 |||| 0.5 |||| 0.5
 +
|- align="center"
 +
| Primer(VR) |||| 0.5 |||| 0.5 |||| 0.5 |||| 0.5 |||| 0.5 |||| 0.5 |||| 0.5 |||| 0.5
 +
|- align="center"
 +
| dH2O || 3.3 || 3.3 || 3.3 || 3.3 || 3.3 || 3.3 || 3.3 || 3.3 || 3.3 || 3.3 || 3.3 || 3.3 || 3.3 || 3.3 || 3.3 || 3.3
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|- align="center"
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| TOTAL || 6 || 6 || 6 || 6 || 6 || 6 || 6 || 6 || 6 || 6 || 6 || 6 || 6 || 6 || 6 || 6
|}
|}
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 +
==Las Check==
 +
 +
22:00 preculture LB-Amp 3 mL
 +
 +
#K084007
 +
#K084012
 +
#T9002
 +
#K091134
 +
#I731012
== Lab Notebook ==
== Lab Notebook ==

Latest revision as of 06:16, 23 September 2009

>Go to the Notebook page

(16_September_2009 <|>18_September_2009)

Contents

Mini Prep

10:30~

  1. I731012(1930bp) pSB1A2(2079bp)
  2. K091134(2048bp) pSB1AK3(3189bp)
  3. S03956(1041bp) pSB1A2(2079bp)
  4. K091118(1985bp) pSB1A2(2079bp)

Digestion Test

14:30~

Double Digestion

volume (uL)
DNA 3 (300ng)
EcoRI 0.20 (4U)
PstI 0.20 (4U)
Buffer(10x) 1
BSA(10x) 1
dW 4.6
total 10



Single Digestion

volume (uL)
DNA 3 (300ng)
EcoRI 0.20 (4U)
Buffer(10x) 1
dW 5.8
total 10


Gel

agar(low electro-osmosis)---0.7%(W)---0.42 g

TAE---60 ml


Energization

135V 27min


Result

from left to right

marker 1 2 3 4(no treatment) marker 1 2 3 4(single digestion) marker 1 2 3 4(double digestion)

ALL:OK

Lux Mutants Sequence

18:30 Mini prep

21:00 PCR

sample number (1) (2) (3) (4) (5) (6) (7) (8) (9) (10) (11) (12) (13) (14) (15) (16)
Big dye v3.1 premix 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2 2
5x sequencing Buffer 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1
LuxR 3 (Mutant number) 1 1
4 1 1
6 1 1
7 1 1
9 1 1
10 1 1
11 1 1
L1 1 1
Primer(VF) 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5
Primer(VR) 0.5 0.5 0.5 0.5 0.5 0.5 0.5 0.5
dH2O 3.3 3.3 3.3 3.3 3.3 3.3 3.3 3.3 3.3 3.3 3.3 3.3 3.3 3.3 3.3 3.3
TOTAL 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6 6

Las Check

22:00 preculture LB-Amp 3 mL

  1. K084007
  2. K084012
  3. T9002
  4. K091134
  5. I731012

Lab Notebook

Chiba-Labwork-14Sep09.jpg Chiba-Labwork-17Sep09-6.jpg Chiba-Labwork-17Sep09-4.jpg Chiba-Labwork-17Sep09-3.jpg

Chiba-Labwork-17Sep09.jpg Chiba-Labwork-17Sep09-5.jpg Chiba-Labwork-17Sep09-2.jpg

Retrieved from "http://2009.igem.org/Team:Chiba/Notebook/Calendar/17_September_2009"