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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
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| BT340 | pCP20 plasmid | LB+Amp LC 30°C,LB+Amp HC 30°C
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| BW23473 | E. coli pir+ strain | LB, 37°C
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| BW23474 | E. coli pir116 strain | LB, 37°C
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| BW25141 | E. coli pir+ strain | LB, 37°C
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| BW25142 | E. coli pir116 strain | LB, 37°C
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| BW5328 /pAH123 | helper plasmid | LB+Amp LC 30°C,LB+Amp HC 30°C
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| MC1061 | E. coli strain for integration | LB, 37°C
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| MG165 | E. coli wil type strain | LB, 37°C
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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
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| I7-1 | 83,3 ng/ul
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| I7-2 | 104 ng/ul
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| I8-1 | 123,8 ng/ul
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| I8-2 | 103,7 ng/ul
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| I9-1 | 117,8 ng/ul
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| I9-2 | 110,4 ng/ul
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| I10-1 | 98,8 ng/ul
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| I10-2 | 93,6 ng/ul
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| PhaP-1 | 29,1 ng/ul
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| PhaP-2 | 13,6 ng/ul
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 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 :)
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