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Lab notes (8/9 - 8/15)

Contents 1 Lab notes (8/9 - 8/15) 1.1 Group: Flagella 1.2 Group: Retinal 1.2.1 Characterization of Beta-carotene 1.2.1.1 Colony PCR on transformants using ninaB fwd and rw primers 1.2.1.2 Harvesting, sonication and UV-vis spektroscopy

Group: Flagella

Group: Retinal

Characterization of Beta-carotene

Start date: 10/8
Methods: ON, sonication, UV-vis spectroscopy
Protocols: CC.1.1[1]

Colony PCR on transformants using ninaB fwd and rw primers

Date: 10/8
Done by: Christian & Tommy
Methods: ON
protocos:

Notes:
ON was made from 4 colonies:
Top 10 - on insert -> OD = 0,008 (100 x diluted)
Top 10 - with K274210 biobrick insert -> OD = 0,011 (100 x diluted)
MG1655 - on insert -> OD = 0,020 (100 x diluted)
Mg1655 - with K274210 biobrick insert -> OD = 0,017 (100 x diluted)
ON was made in 110 ml LB media. colonies with K274210 biobrick insert was grown in media containing ampicillin.
alle the ON cultures was grown for 20 hours at 37 °C.
After 16 hours, ml of the ON colonies was transferred in to 110 LB media and grown for 4 hours to the exponential phase:
Top 10 - on insert -> OD = 0,049 (100 x diluted)
Top 10 - with K274210 biobrick insert -> OD = 0,044 (100 x diluted)
MG1655 - on insert -> OD = 0,007 (100 x diluted)
Mg1655 - with K274210 biobrick insert -> OD = 0,009 (100 x diluted)
Colonies with K274210 biobrick insert was grown in media containing ampicillin.

Harvesting, sonication and UV-vis spektroscopy

Date: 11/8
Done by: Christian & Tommy
Methods: Cell harvesting, sonication and UV-vis messurements
protocos: none

Notes:
100 mL Cell cultures was centrifuged for 5 min at 14000 RPM. the supernantant was discarded and cells were resuspended in 5 mL acetone (99,9%), except the top 10 with the K274210 biobrick insert wicht was resuspended in 10 mL acetone (suorce of error.). The resuspended cells were sonicated for 5 min. the samples were spun down and the supernantant were transferrede to new tubes, cell bebri was discarded.

A standart curve was made from pure Beta-carotene,