Team:SDU-Denmark/labnotes5
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== Group: Flagella == | == Group: Flagella == | ||
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+ | == Group: Retinal == | ||
+ | === Characterization of Beta-carotene === | ||
+ | Start date: 10/8<br> | ||
+ | Methods: ON, sonication, UV-vis spectroscopy<br> | ||
+ | Protocols: CC.1.1[https://2010.igem.org/Team:SDU-Denmark/protocols#CP1.1] | ||
+ | ==== Colony PCR on transformants using ninaB fwd and rw primers ==== | ||
+ | Date: 10/8<br> | ||
+ | Done by: Christian & Tommy<br> | ||
+ | Methods: ON<br> | ||
+ | protocos: | ||
+ | <br><br> | ||
+ | Notes:<br> | ||
+ | ON was made from 4 colonies:<br> | ||
+ | Top 10 - on insert -> OD = 0,008 (100 x diluted)<br> | ||
+ | Top 10 - with K274210 biobrick insert -> OD = 0,011 (100 x diluted)<br> | ||
+ | MG1655 - on insert -> OD = 0,020 (100 x diluted)<br> | ||
+ | Mg1655 - with K274210 biobrick insert -> OD = 0,017 (100 x diluted)<br> | ||
+ | ON was made in 110 ml LB media. colonies with K274210 biobrick insert was grown in media containing ampicillin.<br> | ||
+ | alle the ON cultures was grown for 20 hours at 37 °C.<br> | ||
+ | After 16 hours, ml of the ON colonies was transferred in to 110 LB media and grown for 4 hours to the exponential phase:<br> | ||
+ | Top 10 - on insert -> OD = 0,049 (100 x diluted)<br> | ||
+ | Top 10 - with K274210 biobrick insert -> OD = 0,044 (100 x diluted)<br> | ||
+ | MG1655 - on insert -> OD = 0,007 (100 x diluted)<br> | ||
+ | Mg1655 - with K274210 biobrick insert -> OD = 0,009 (100 x diluted)<br> | ||
+ | Colonies with K274210 biobrick insert was grown in media containing ampicillin.<br> | ||
+ | |||
+ | ==== Harvesting, sonication and UV-vis spektroscopy ==== | ||
+ | Date: 11/8<br> | ||
+ | Done by: Christian & Tommy<br> | ||
+ | Methods: Cell harvesting, sonication and UV-vis messurements<br> | ||
+ | protocos: none<br> | ||
+ | <br> | ||
+ | Notes:<br> | ||
+ | 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.<br> | ||
+ | A standart curve was made from pure Beta-carotene,<br> |
Revision as of 11:33, 17 August 2010