Team:UNIPV-Pavia/Calendar/July/settimana1

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Revision as of 11:14, 5 July 2010

JULY: WEEK 1

June, 28th

Inoculum from glycerol stock for I7-1, I7-2, I8-1, I8-2, I9-1, I9-2, I10-1 and I10-2 in 5ml LB+Amp. Cultures were gown ON at 37°C 220rpm.

Inoculum of PhaP-1 and PhaP-2 in 5ml LB+Kan and ON growth at 37°C 220rpm.

We received strains and plasmid from ???Yale University???

name	description	growth conditions
BT340	pCP20 plasmid	LB+Amp LC 30°C,LB+Amp HC 30°C
BW23473	E. coli pir+ strain	LB, 37°C
BW23474	E. coli pir116 strain	LB, 37°C
BW25141	E. coli pir+ strain	LB, 37°C
BW25142	E. coli pir116 strain	LB, 37°C
BW5328 /pAH123	helper plasmid	LB+Amp LC 30°C,LB+Amp HC 30°C
MC1061	E. coli strain for integration	LB, 37°C
MG165	E. coli wil type strain	LB, 37°C

June, 29th

All cultures were grown. From I7-1, I7-2, I8-1, I8-2, I9-1, I9-2, I10-1 and I10-2 5ul were aliquoted and diluted in 500ul LB+Amp for a preliminary TECAN test. Cultures were MiniPrepped with the following quantifications (NanoDrop):

came	quantification
I7-1	83,3 ng/ul
I7-2	104 ng/ul
I8-1	123,8 ng/ul
I8-2	103,7 ng/ul
I9-1	117,8 ng/ul
I9-2	110,4 ng/ul
I10-1	98,8 ng/ul
I10-2	93,6 ng/ul
PhaP-1	29,1 ng/ul
PhaP-2	13,6 ng/ul

Screening of I7, I8, I9, I10

All strains received were on blotting paper disks, that were placed on an LB agar plates (with/without antiobiotic, see growth conditions), resuspended wih 80ul LB and then streaked. After plate streaking, disks were trasnferred in falcon containing liquid LB.

June, 30th

July, 1st

July, 2nd

Screening of new colonies of I7 and I8. New screening for I9, I10

This time all the clones are OK :)

week 1

@@ Line 43: / Line 43: @@
 ==June, 29th==
-All cultures were grown. From each culture, 5ul were aliquoted and diluted in 500ul LB+Amp for a preliminary TECAN test.
+All cultures were grown. From I7-1, I7-2, I8-1, I8-2, I9-1, I9-2, I10-1 and I10-2 5ul were aliquoted and diluted in 500ul LB+Amp for a preliminary TECAN test.
+Cultures were MiniPrepped with the following quantifications (NanoDrop):
+{| border='1' align='center'
+|| '''came''' ||'''quantification'''
+|-
+|| I7-1 || 83,3 ng/ul
+|-
+|| I7-2 || 104 ng/ul
+|-
+|| I8-1 || 123,8 ng/ul
+|-
+|| I8-2 ||  103,7 ng/ul
+|-
+|| I9-1 || 117,8 ng/ul
+|-
+|| I9-2 || 110,4 ng/ul
+|-
+|| I10-1 || 98,8 ng/ul
+|-
+|| I10-2 || 93,6 ng/ul
+|-
+|| PhaP-1 || 29,1 ng/ul
+|-
+|| PhaP-2 || 13,6 ng/ul
+|}
 [[Image:Unipv_screening_I7_I8_I9_I10.jpg|300px|thumb|center|Screening of I7, I8, I9, I10]]