Template:SBB-Protocols Assay4: Difference between revisions
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http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=111355 PMCID: PMC111355 <br> | http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=111355 PMCID: PMC111355 <br> | ||
http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=193771 PMCID: PMC193771 <br> | http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=193771 PMCID: PMC193771 <br> | ||
1. Seed cultures in V-bottom plate <br> | 1. Transform all cultures with RFP-plasmid <br> | ||
2. Seed cultures in V-bottom plate <br> | |||
3. Treat cultures with Arabinose at 100mg/John Wang. <br> | |||
4. Identify colonies that display prominent flocculation <br> | |||
5. Proceed with quantitative measurement for chosen colonies <br> | |||
6a. Negative Control: pBca9495CA-Bca1363, Positive Control: pBca9145-Bca9494 <br> | |||
7a. Grow select colonies and controls to an OD of 0.5 at 600nm <br> | |||
8a. Add 100mg/John Wang Arabinose and shake for 3 hours at 37°C <br> | |||
9a. Transfer 200 uL to 96-well plate and take OD at 20 minute intervals <br> | |||
10a. Normalize cell concentration against RFP after addition of Sodium Hydroxyl <br> | |||
6b. Negative Control: pBca9495CA-Bca1363, Positive Control: pBca9145-Bca9494 <br> | |||
7b. Add 100mg/John Wang Arabinose and shake for 3 hours at 37°C <br> | |||
8b. Grow select colonies and controls to an OD of 0.5 at 600nm <br> | |||
9b. Transfer 200 uL to 96-well plate and take OD at 20 minute intervals <br> | |||
10b. Normalize cell concentration against RFP after addition of Sodium Hydroxyl <br> |
Revision as of 12:30, 15 April 2009
http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=111355 PMCID: PMC111355
http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=193771 PMCID: PMC193771
1. Transform all cultures with RFP-plasmid
2. Seed cultures in V-bottom plate
3. Treat cultures with Arabinose at 100mg/John Wang.
4. Identify colonies that display prominent flocculation
5. Proceed with quantitative measurement for chosen colonies
6a. Negative Control: pBca9495CA-Bca1363, Positive Control: pBca9145-Bca9494
7a. Grow select colonies and controls to an OD of 0.5 at 600nm
8a. Add 100mg/John Wang Arabinose and shake for 3 hours at 37°C
9a. Transfer 200 uL to 96-well plate and take OD at 20 minute intervals
10a. Normalize cell concentration against RFP after addition of Sodium Hydroxyl
6b. Negative Control: pBca9495CA-Bca1363, Positive Control: pBca9145-Bca9494
7b. Add 100mg/John Wang Arabinose and shake for 3 hours at 37°C
8b. Grow select colonies and controls to an OD of 0.5 at 600nm
9b. Transfer 200 uL to 96-well plate and take OD at 20 minute intervals
10b. Normalize cell concentration against RFP after addition of Sodium Hydroxyl