Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Experiment Videos

Probe generation directly from small numbers of cells for DNA microarray studies.

C C Xiang1, M Chen, O A Kozhich

  • 1NIMH/NIH, Bethesda, MD, USA.

Biotechniques
|March 5, 2003
PubMed
Summary

This study introduces a new RNA amplification protocol enabling gene expression profiling from minimal cell samples. The method is reproducible and yields results comparable to existing techniques, expanding applications in cell analysis.

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

[Epidemiological investigation of tinnitus in Sichuan and Chongqing].

Zhonghua er bi yan hou tou jing wai ke za zhi = Chinese journal of otorhinolaryngology head and neck surgery·2021
Same author

Characterisation of insulin-resistant phenotype of cultured rat primary adipose cells.

Diabetologia·2007
Same author

Gene expression profiling of human placentas from preeclamptic and normotensive pregnancies.

Molecular human reproduction·2006
Same author

cDNA microarray technology and its applications.

Biotechnology advances·2003
Same author

Oncogenic c-Ki-ras but not oncogenic c-Ha-ras up-regulates CEA expression and disrupts basolateral polarity in colon epithelial cells.

The Journal of biological chemistry·1997
Same author

Transient gene expression from yeast artificial chromosome DNA in mammalian cells is enhanced by adenovirus.

Nucleic acids research·1997

Area of Science:

  • Molecular Biology
  • Genomics
  • Biotechnology

Background:

  • Previous RNA expression profiling methods required significant RNA input.
  • Existing techniques were not suitable for limited cell samples obtained via needle biopsy or cell sorting.

Purpose of the Study:

  • To develop a novel RNA amplification protocol for gene expression profiling from minimal cell samples (approx. 10 cells).
  • To enable reliable and reproducible expression profiling from challenging sample types.

Main Methods:

  • A multi-cycle RNA amplification protocol using random 9-mers and specific RNA polymerase recognition sequences (T3 and T7).
  • Direct cell lysis into reverse transcription buffer followed by three rounds of RNA amplification.
  • Preparation of fluorescent probes from amplified RNA for expression profiling using 11,232-element arrays.

Related Experiment Videos

Main Results:

  • The new protocol successfully amplified RNA from approximately 10 cells.
  • Expression profiles generated from amplified RNA were highly reproducible and reliable.
  • Results were comparable to those obtained using the RiboAmp RNA kit and unamplified templates.

Conclusions:

  • The developed RNA amplification protocol significantly expands the utility of gene expression profiling for limited cell samples.
  • This method offers a reliable and reproducible alternative for analyzing gene expression in challenging biological samples.