Revision as of 15:33, 9 August 2013

Notebook : August 8

Damir, Nadia

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Expected sizes :

We obtained fragments of the right size. Now we can purify the PSB3K3 plasmid.

2.1 - Migration of the remaining 45µL of BBa_J004450 digested by EcoRI/Pst1

Anaïs

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Now we can do a gel purification of the highest band which contains the PSB3K3 plasmid.

2.2 - Electroelution of the highest band to extract the PSB3K3 plasmid from the gel

Nadia

We let the plasmid precipitate during the night.

Anaïs

Colony counting :
- Low concentration petri dish : 47 colonies
- High concentration petri dish : 145 colonies

@@ Line 5: / Line 5: @@
 =='''Lab work'''==
-==='''Aerobic/Anaerobic regulation system'''===
+==='''A - Aerobic/Anaerobic regulation system'''===
-===='''Preparation of the PSB3K3 backbone plasmid'''====
+===='''Obtaining the PSB3K3 backbone plasmid'''====
 =====1 - Electrophoresis of BBa_J004450 digested by EcoRI/Pst1 to check if the digestion was right=====
@@ Line 50: / Line 50: @@
   We let the plasmid precipitate during the night.
+===='''Obtaining RBS_LacZ+Term_PSB1C3'''====
+=====1 - Colony PCR on e.coli with RBS_LacZ+Term_PSB1C3 for 25 colonies=====
+Anaïs
+*Colony counting :
+**Low concentration petri dish : 47 colonies
+**High concentration petri dish : 145 colonies
+*Preparation of 700µL of Master mix
+**H<sub>2</sub>O : 590µL
+**dNTP : 28µL
+**VF2 primer : 3.5µL
+**VR primer : 3.5µL
+**DreamTaq buffer 10x : 70µL
+**DreamTaq enzyme : 5µL
 {{Team:Paris_Saclay/incl_fin}}