Team:Chiba/Notebook/Calendar/17 September 2009

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(Difference between revisions)
(→Lux Mutants Sequence)
(→Lux Mutants Sequence)
Line 90: Line 90:
| 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"
-
| PstI || 0.20 (4U)
+
| LuxR 3 (Mutant number) || 1 || 1 ||
|- align="center"
|- align="center"
-
| Buffer(10x) || 1
+
| 4 |||||| 1 || 1
|- align="center"
|- align="center"
-
| BSA(10x)|| 1
+
| 6 |||||||||| 1 || 1 ||
|- align="center"
|- align="center"
-
| dW || 4.6
+
| 7 |||||||||||||| 1 || 1
|- align="center"
|- align="center"
-
| total || 10
+
| 9 |||||||||||||||||| 1 || 1
 +
|- align="center"
 +
| 10 |||||||||||||||||||||| 1 || 1
 +
|- align="center"
 +
| 11 |||||||||||||||||||||||||| 1 || 1
 +
|- align="center"
 +
| L1 |||||||||||||||||||||||||||||| 1 || 1
 +
|- align="center"
 +
| 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
 +
|- align="center"
 +
| TOTAL || 6 || 6 || 6 || 6 || 6 || 6 || 6 || 6 || 6 || 6 || 6 || 6 || 6 || 6 || 6 || 6
|}
|}

Revision as of 06:07, 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

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

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