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Isolation and Characterization of Intact Phycobilisome in Cyanobacteria

3.4K Views

06:26 min

November 10th, 2021

November 10th, 2021

3,449 Views

0:04

Introduction

0:38

Cell Culture and Harvesting

1:13

Cells Lysis

2:43

Preparation of the Sucrose Gradient Buffers and PBS Isolation

3:32

Measurement of PBS Fluorescence at 77K

4:00

Results: Spectral Evaluation of Phycobilisomes

5:50

Conclusion

文字起こし

It is a simple and a fast protocol that allow people unfamiliar with cyanobacteria to isolate phycobilisome at ease. The advantage of this technique is that it is adapted to a small scale and it is easy to perform. It is a simple, reliable, and ef

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The current protocol details the isolation of phycobilisomes from cyanobacteria by centrifugation through a discontinuous sucrose density gradient. The fractions of intact phycobilisomes are confirmed by 77K fluorescent emission spectrum and SDS-PAGE analysis. The resulting phycobilisome fractions are suitable for negative staining of TEM and mass spectrometry analysis.

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