Team:Paris/August 20

From 2008.igem.org

(Difference between revisions)
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The plates obtained from the speading of yesterday can't be used because there are not single colonies.
The plates obtained from the speading of yesterday can't be used because there are not single colonies.
<br>We have to try again, but with a stronger dilution of the bacteria or with a smaller volume of spreading.
<br>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===
 +
[[Team:Paris/Notebook/Protocols#Concentration of the Miniprep|standard protocol]]
 +
 +
{| border="1" style="text-align: center"
 +
|'''Miniprep'''
 +
|'''Biobrick'''
 +
|'''Concentration (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===
 +
 +
to be modified
 +
 +
[[Team:Paris/Notebook/Protocols#Digestion|Protocol Digestion]]
 +
 +
{| border="1" style="text-align: center"
 +
|'''Plasmid'''
 +
|'''Description'''
 +
|'''Miniprep used'''
 +
|'''Enzymes'''
 +
|-
 +
|MP3.3 ?
 +
|B0015 (double terminator B0010-B0012) - FV
 +
|4
 +
|EcoRI and XbaI
 +
|-
 +
|MP119
 +
|promoter J23101 - BV
 +
|1
 +
|SpeI and PstI
 +
|-
 +
|MP104
 +
|PTet (Tet promoter) - BV
 +
|1
 +
|SpeI and PstI
 +
|-
 +
|MP114
 +
|TetR - FI
 +
|1
 +
|EcoRI and SpeI
 +
|-
 +
|MP143
 +
|gfp generator - BI
 +
|2
 +
|SpeI and PstI
 +
|}

Revision as of 16:02, 20 August 2008

← Yesterday

↓ Calendar ↑

Tomorrow →

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 Concentration (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

to be modified

Protocol Digestion

Plasmid Description Miniprep used Enzymes
MP3.3 ? B0015 (double terminator B0010-B0012) - FV 4 EcoRI and XbaI
MP119 promoter J23101 - BV 1 SpeI and PstI
MP104 PTet (Tet promoter) - BV 1 SpeI and PstI
MP114 TetR - FI 1 EcoRI and SpeI
MP143 gfp generator - BI 2 SpeI and PstI
Retrieved from "http://2008.igem.org/Team:Paris/August_20"