Team:Paris/July 19

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Revision as of 13:44, 21 July 2008

MiniPrep

The protocol of MiniPrep of Quiagen has been done on all the clones cultivated on the 18th. J23109 clones 1,2,5
E0030 clones 1,2,3
pSB1A3 clones 1,2,3
C0179 clones 1,2,7
C0079 clones 1,2,3
R0079 clones 3,4,6
E1010 clones 1,2,3
B0034 clones 1,2,3
J23101 clones 1,2,3
E0040 clones 1,2
R0040 clones 1,2

We did an error during the manipulation (probably an exchange between some tubes) so we decided to screen again clones of B0034 and R0040.

Quantitative analysis of ADN

gel 1

Biobricks	Clone	Expected size	Measured size
E1010	1	5.106 kb	~6-8 kb
E1010	2	5.106 kb	~6-8 kb
C0079	1	5.181 kb	~6-8 kb
C0079	2	5.181 kb	~6-8 kb
C0079	3	5.181 kb	~6-8 kb
J23101 : problem !!!	1	3.037 kb	~6-8 kb
J23101 : problem !!!	2	3.037 kb	~6-8 kb
J23101 : problem !!!	3	3.037 kb	~6-8 kb
B0034	2	2.091 kb	~2 kb
B0034	X	2.091 kb	~2 kb
B0034 : problem !!!	Y	2.091 kb	~5 kb
E0040	1	2.799 kb	~2.5-3 kb
E0040	2	2.799 kb	~2.5-3 kb
R0040	?	2.133 kb	~2 kb

We see bad results for B0034 (bands 10 to 12), R0040 (band 15). We'll try again!

gel 2

We see bad results for J23109 (bands 1 to 3), C0179 (bands 14 to 16). We'll try again!

Screening R0040 and B0034

PCR

For 1 tube : 25µL Quick Load
1µL Oligo F
1µL Oligo R
23µL H₂O

Programm SCREENIN

Electrophoresis

Band 0 : ladder 100pb Band 2 : B0034 clone 1 -> no result Band 3 : B0034 clone 2 -> ok Band 4 : R0040 clone 1 -> ok Band 5 : R0040 clone 2 -> problem Band 6 : R0040 ? -> problem

@@ Line 116: / Line 116: @@
 Programm SCREENIN
-===
+===Electrophoresis===
+Band 0 : ladder 100pb
+Band 2 : B0034 clone 1 -> no result
+Band 3 : B0034 clone 2 -> ok
+Band 4 : R0040 clone 1 -> ok
+Band 5 : R0040 clone 2 -> problem
+Band 6 : R0040 ? -> problem