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Measuring Gene Expression in Bombarded Barley Aleurone Layers with Increased Throughput
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Genetics
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JoVE Journal Genetics
Measuring Gene Expression in Bombarded Barley Aleurone Layers with Increased Throughput

Measuring Gene Expression in Bombarded Barley Aleurone Layers with Increased Throughput

DOI:
10.3791/56728-v

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10:29 min

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March 30, 2018

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, , , ,
1Department of Biology,Colby College

Chapters

  • 00:05Title
  • 00:44Preparation of Barley Grains and Plasmid DNA for Bombardment
  • 02:42Particle Bombardment of Embryoless Barley Grains
  • 04:42Extraction of Soluble Protein from the Bombarded Grains
  • 06:24Measurement of GUS Activity from Grain Extracts
  • 08:15Results: Measurement of Gene Expression in Barley Aleurone Cells after Particle Bombardment
  • 09:50Conclusion

Summary

Automatic Translation

Automatic Translation

An improved protocol is presented for the measurement of transient gene expression from reporter constructs in barley aleurone cells after particle bombardment. The combination of automated grain grinding with 96-well plate enzyme assays provides high throughput for the procedure.

Tags

Keywords: Gene ExpressionBarley AleuroneParticle BombardmentReporter ConstructsAbscisic AcidHigh-throughputSeed Coat PeelingVermiculiteUBI LuciferaseGUS ReporterEffector PlasmidMicrocarriersParticle Delivery SystemRupture Disk

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