Team:Paris/August 20

From 2008.igem.org

(Difference between revisions)
(→Measurement of concentration of minipreps)
(→Digestion)
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===Digestion===
===Digestion===
-
to be modified
 
[[Team:Paris/Notebook/Protocols#Digestion|Protocol Digestion]]
[[Team:Paris/Notebook/Protocols#Digestion|Protocol Digestion]]
{| border="1" style="text-align: center"
{| border="1" style="text-align: center"
-
|'''Plasmid'''
+
|'''Name'''
 +
|'''Miniprep used'''
|'''Description'''
|'''Description'''
-
|'''Miniprep used'''
+
|'''Vol MP (µl)'''
 +
|'''Vol H2O (µl)'''
|'''Enzymes'''
|'''Enzymes'''
|-
|-
-
|MP3.3 ?
+
|D166
-
|B0015 (double terminator B0010-B0012) - FV
+
|MP165.1
-
|4
+
|- FV
 +
|7.63
 +
|17
|EcoRI and XbaI
|EcoRI and XbaI
|-
|-
-
|MP119
+
|D167
-
|promoter J23101 - BV
+
|MP166.1
-
|1
+
| - BV
-
|SpeI and PstI
+
|8.9
-
|-
+
|15.8
-
|MP104
+
|EcoRI and XbaI
-
|PTet (Tet promoter) - BV
+
-
|1
+
-
|SpeI and PstI
+
-
|-
+
-
|MP114
+
-
|TetR - FI
+
-
|1
+
-
|EcoRI and SpeI
+
|-
|-
-
|MP143
+
|D131
-
|gfp generator - BI
+
|MP122.2
-
|2
+
|- BV
-
|SpeI and PstI
+
|10.2
 +
|14.5
 +
|XbaI and PstI
|}
|}

Revision as of 16:11, 20 August 2008

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Contents

Screening of the cloning of E0240 and FlhDC+promotor

Spreading the clones in order to obtain single colonies

Strain Resistance Ligation DNA cloned vector expected size of the fragment amplified by VF & VR mesured size
S159.1 kanamycine L139.1 E0240 (GFP tripart) pSB3K3 1192 bp 1,5 kb
1,1 kb
0,6 kb
S161.1 ampicilline L142.7 FlhDC+promotor pSB1A2 1403 bp 1,4
0,4 kb
0,3 kb

The plates obtained from the speading of yesterday can't be used because there are not single colonies.
We have to try again, but with a stronger dilution of the bacteria or with a smaller volume of spreading.


Digestion

Aim : Construction of "pFlgA-RBS-YFP-dbl ter"

Measurement of concentration of minipreps

standard protocol

Miniprep Biobrick C° (µg/mL) ratio 260/280
MP164.1 E0422 95 1.69
MP164.2 E422 90 1.76
MP165.1 E0430 131 1.74
MP165.2 E0430 nd nd
MP166.1 E0432 112 1.65
MP166.2 E0432 79 1.6
MP167.1 E0420 199 1.72
MP167.2 E0420 194 1.73
MP168.1 I732078 111 1.65
MP168.2 I732078 104 1.67
MP122.1 E0840 56 1.58
MP122.2 E0840 98 1.63

Digestion

Protocol Digestion

Name Miniprep used Description Vol MP (µl) Vol H2O (µl) Enzymes
D166 MP165.1
7.63 17 EcoRI and XbaI
D167 MP166.1 - BV 8.9 15.8 EcoRI and XbaI
D131 MP122.2
10.2 14.5 XbaI and PstI
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