Experiment Overview
| Repository ID: | FR-FCM-ZY88 | Experiment name: | JN27 vs JN27.SH binding | MIFlowCyt score: | 90.00% |
| Primary researcher: | Jennifer Soundy | PI/manager: | Darren Day | Uploaded by: | Jennifer Soundy |
| Experiment dates: | 2016-06-08 - 2016-06-09 | Dataset uploaded: | Jul 2017 | Last updated: | Jul 2017 |
| Keywords: | [Bacteria] [Aptamer binding] [Pseudomonas aeruginosa] [SELEX progression] [FAM labelled aptamer] | Manuscripts: | |||
| Organizations: |
Victoria University of Wellington, School of Biological Sciences, Wellington, Wellington (New Zealand)
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| Purpose: | The purpose of this experiment was to test our hypothesis that truncation of the aptamer candidates to just a stem-loop structure would increase the binding seen compared to the full length aptamer. | ||||
| Conclusion: | We found that our hypothesis was true and truncation of the JN27 aptamer to the JN27.SH aptamer, removing the non essential sequence that did not comprise the stem loop motif, made the aptamer bind better than before. This is seen as a right shift in the FITC fluorescence compared between the two aptamers at the same concentration. | ||||
| Comments: | The experiment was done with 4 different concentrations of each aptamer. 50 nM, 100 nM, 250 nM and 500 nM | ||||
| Funding: | Not disclosed | ||||
| Quality control: | The same flow cytometer and settings are used for all experiments. The PBS used as flow buffer to re-suspend the bacteria in was run on its own to check for contamination or particulate that shows up as the same size as our target bacteria. | ||||
Experiment variables
| Conditions | |
|---|---|
| · JN27 aptamer | JN27_100_005.fcs · JN27_250_007.fcs · JN27_500_009.fcs · JN27_50_003.fcs |
| · JN27.SH aptamer | JN27_100 short_006.fcs · JN27_250 short_008.fcs · JN27_50 short_004.fcs · JN27_500 short_010.fcs |
| Doses | |
|---|---|
| · 50 nM | JN27_50 short_004.fcs · JN27_50_003.fcs |
| · 100 nM | JN27_100 short_006.fcs · JN27_100_005.fcs |
| · 250 nM | JN27_250 short_008.fcs · JN27_250_007.fcs |
| · 500 nM | JN27_500 short_010.fcs · JN27_500_009.fcs |
| · None | Control cells_PA692_002.fcs · Control cells_PBS_001.fcs |
