Difference between revisions of "Part:BBa K523002:Experience"
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how you used this part and how it worked out. | how you used this part and how it worked out. | ||
| − | + | ==Applications of BBa_K523002== | |
| + | =UMA_MALAGA 2022 Improvement= | ||
| + | This year we decided to improve BBa_K523002 by deleting the native RBS and optimizing the whole sequence. We used our new part (<partinfo>BBa_K4368002</partinfo>) to have an enzymatic complex named CAZymes with cenA and cex to degrade cellulose into glucose. | ||
| + | |||
| + | ===Characterization=== | ||
| + | The expression cassette sequence was digested with EcoRI and PstI enzymes and subsequently ligated with a chloramphenicol resistant plasmid backbone (Cm). | ||
| + | Transforming bacteria were created with this plasmid and seeded on LB-Agar+Cm plates. After growth, colonies were selected based on their color (white) and DNA extraction was performed using the Promega PureYield Plasmid Miniprep System kit. | ||
| + | The resulting DNA is used for further digestion with EcoRI and PstI. The digests are then run on a 0.75% agarose gel at 90 mV voltage and constant amperage. BioRad brand RedSafe is used as an intercalating developing agent. | ||
| + | |||
| + | ===Enzyme digestion=== | ||
| + | [[File:bglX.png|350px|center|]] | ||
| + | |||
| + | ===Conclusion=== | ||
| + | We succeded in the selection of transformed bacteria with this gene. We observed degradation of celulose in plates using the red congo assay and measuring the hidrolisis halo. | ||
| + | |||
| + | *'''Group:''' [https://2022.igem.wiki/uma-malaga/index.html UMA_MALAGA] | ||
| + | *'''Author:''' Molina Calvo, Alonso | ||
===User Reviews=== | ===User Reviews=== | ||
<!-- DON'T DELETE --><partinfo>BBa_K523002 StartReviews</partinfo> | <!-- DON'T DELETE --><partinfo>BBa_K523002 StartReviews</partinfo> | ||
Latest revision as of 10:51, 13 October 2022
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Please enter
how you used this part and how it worked out.
Applications of BBa_K523002
UMA_MALAGA 2022 Improvement
This year we decided to improve BBa_K523002 by deleting the native RBS and optimizing the whole sequence. We used our new part (BBa_K4368002) to have an enzymatic complex named CAZymes with cenA and cex to degrade cellulose into glucose.
Characterization
The expression cassette sequence was digested with EcoRI and PstI enzymes and subsequently ligated with a chloramphenicol resistant plasmid backbone (Cm). Transforming bacteria were created with this plasmid and seeded on LB-Agar+Cm plates. After growth, colonies were selected based on their color (white) and DNA extraction was performed using the Promega PureYield Plasmid Miniprep System kit. The resulting DNA is used for further digestion with EcoRI and PstI. The digests are then run on a 0.75% agarose gel at 90 mV voltage and constant amperage. BioRad brand RedSafe is used as an intercalating developing agent.
Enzyme digestion
Conclusion
We succeded in the selection of transformed bacteria with this gene. We observed degradation of celulose in plates using the red congo assay and measuring the hidrolisis halo.
- Group: UMA_MALAGA
- Author: Molina Calvo, Alonso
User Reviews
UNIQ9a2dcdc1293a8d22-partinfo-00000001-QINU UNIQ9a2dcdc1293a8d22-partinfo-00000002-QINU