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Single Cell Multiplex Reverse Transcription Polymerase Chain Reaction After Patch-clamp
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Neuroscienze
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JoVE Journal Neuroscienze
Single Cell Multiplex Reverse Transcription Polymerase Chain Reaction After Patch-clamp

Single Cell Multiplex Reverse Transcription Polymerase Chain Reaction After Patch-clamp

DOI:
10.3791/57627-v

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10:44 min

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June 20, 2018

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, , ,
1UPMC Univ Paris 06, INSERM, CNRS, Neuroscience Paris Seine – Institut de Biologie Paris Seine (NPS – IBPS),Sorbonne Université

Capitoli

  • 00:04Titolo
  • 00:57Acute Slice Preparation
  • 02:53Single Cell RT-PCR after Patch-clamp Recording
  • 05:54PCR Validation
  • 08:47Results: Characterization of Layer V Neocortical Pyramidal Cells
  • 09:57Conclusion

Summary

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This protocol describes the critical steps and precautions required to perform single cell multiplex reverse transcription polymerase chain reaction after patch-clamp. This technique is a simple and effective method to analyze the expression profile of a predetermined set of genes from a single cell characterized by patch-clamp recordings.

Tags

Keywords: Single CellMultiplexReverse TranscriptionPolymerase Chain ReactionPatch-clampNeuroscienzeNeuronal PopulationTransmission SystemsGene ExpressionAstrocytesAnimal ModelsBrain DissectionVibratome SectioningWhole-cell RecordingCytoplasm HarvestingRT-PCRGDNA Contamination

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