Difference between revisions of "Part:BBa K5097015:Experience"
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Team Oneonta 2024 used this part to purify RFP for studies on the effect of pH on the emission of this fluorescent protein. We expressed composite parts BBa_K5097016 and BBa_K5097017 in E. coli DH5alpha cells, and lysed the cells by treating with lysozyme and sonication. Proteins were purified via Ni-IMAC chromatography and eluted in 50mM sodium phosphate, 300mM sodium chloride, and 500mM imidazole at pH8.0. Proteins were concentrated and exchanged into PBS via centrifugal diafiltration. | Team Oneonta 2024 used this part to purify RFP for studies on the effect of pH on the emission of this fluorescent protein. We expressed composite parts BBa_K5097016 and BBa_K5097017 in E. coli DH5alpha cells, and lysed the cells by treating with lysozyme and sonication. Proteins were purified via Ni-IMAC chromatography and eluted in 50mM sodium phosphate, 300mM sodium chloride, and 500mM imidazole at pH8.0. Proteins were concentrated and exchanged into PBS via centrifugal diafiltration. | ||
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| + | :::::::::Figure 1: Coomasie stained PAGE gel showing purification of BFP-6XHis and RFP-6XHis | ||
===User Reviews=== | ===User Reviews=== | ||
Latest revision as of 18:03, 30 September 2024
This experience page is provided so that any user may enter their experience using this part.
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how you used this part and how it worked out.
Applications of BBa_K5097015
Team Oneonta 2024 used this part to purify RFP for studies on the effect of pH on the emission of this fluorescent protein. We expressed composite parts BBa_K5097016 and BBa_K5097017 in E. coli DH5alpha cells, and lysed the cells by treating with lysozyme and sonication. Proteins were purified via Ni-IMAC chromatography and eluted in 50mM sodium phosphate, 300mM sodium chloride, and 500mM imidazole at pH8.0. Proteins were concentrated and exchanged into PBS via centrifugal diafiltration.
- Figure 1: Coomasie stained PAGE gel showing purification of BFP-6XHis and RFP-6XHis
User Reviews
UNIQce14750a77245d7a-partinfo-00000000-QINU UNIQce14750a77245d7a-partinfo-00000001-QINU