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Polysome Profiling in Leishmania, Human Cells and Mouse Testis
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Biochemistry
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JoVE Journal Biochemistry
Polysome Profiling in Leishmania, Human Cells and Mouse Testis
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Polysome Profiling in Leishmania, Human Cells and Mouse Testis

DOI:
10.3791/57600-v

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14:32 min

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April 08, 2018

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, , , , , , , , ,
1Department of Cell Biology and Biochemistry,Texas Tech University Health Sciences Center, 2Department of Biological Sciences,Texas Tech University, 3CISER (Center for the Integration of STEM Education & Research),Texas Tech University

Chapters

  • 00:00Title
  • 01:24Lysate Preparation
  • 03:51Cytoplamic Lysate Preparation from Cultured Human Cells
  • 05:15Cytoplasmic Lysate Preparation from Mouse Testis
  • 06:53Sucrose Gradient Preparation and Ultracentrifugation
  • 09:21Polysome Fractionation and Sample Collection
  • 11:45Analysis of the Fractions
  • 12:27Results
  • 14:14Conclusions

Summary

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The overall goal of polysome profiling technique is analysis of translational activity of individual mRNAs or transcriptome mRNAs during protein synthesis. The method is important for studies of protein synthesis regulation, translation activation and repression in health and multiple human diseases.

Tags

Polysome ProfilingMRNA TranslationProtein Synthesis RegulationLeishmaniaHuman CellsMouse TestisSucrose GradientUltracentrifugationRibosome FractionationCytoplasmic LysateCycloheximideCell CulturePBS WashCell Counting

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