University:
Swansea University
Course:
PM134 | Microbiology
Academic year:

2023
Views:

299

Pages:

2
Author:

Sanjeev K

EXPERIMENT NO:9 DATE: 812-2022 MOLECULAR EXCLUSION CHROMATOGRAPH AIM to differentiate hibstances wing molecular exclusion chromatography PRINCIPLE Molecular exclusion chromatography, also knour If thatior as and porrmeation cheomatography or gel fil chmamatography. It is a techniques in which mdeules in solution are separated by thein ze a molecular weight. A mixture liquid molecules durch is apptrea to a chromatography colum whien contains a solid scope sphorus, or the fond og ma in the "beads". The beads au as traps or sieves and function do filter smallt whien berorrle temporarile Large sample molentes Op trapped molecules within the pores partially penetrate the pores, where as smaller molecules cam alless mest for au pores. in Loage modevils elute forest, smaller molecule elute later, cwhile molecules which are very small puell elity last. Particles f different sizes cuill Elite through la stationary phall as differem rates MATERIALS REQUIRED cmomatography column, beaker glau wool, wool, Sephadex pss buffer, meanglene bure, cytbumsome c PROCEDURG * Take a clean chromatography column and add myficient glass wool. * fill the column with Sephadex keeping in mind that there is no leakage. * Ares a feus minutes, add PBs buffer * Open the topcok al colled the eluent in a bea my buffer ker where it is made wire that ang Melnykhe blue, cytochrome C and Pas buka were added to the colurmay the components wore allowed to separate RESULT Cytocmome C eluted fort while Methylene blue was ented after that. The naicates the the molecular beef cytochrome & was greater than cize Meputene blue