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Cell Culture Mycoplasma Detection by PCR

HP Huan Pang

Published: Apr 20, 2012 DOI: 10.21769/BioProtoc.207 Views: 19937

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Abstract

DNA was extracted from the supernatant of each sample. After PCR-amplification of mycoplasma DNA, detection was performed by gel electrophresis. The PCR primers were designed to cover the consensus sequences that can detect all types of mycoplasma species.

Materials and Reagents

  1. Ampli Taq Gold (Life Technologies, InvitrogenTM, catalog number: 4338856 )
  2. Sodium acetate
  3. Ethanol
  4. Phenol
  5. MgCl2
  6. PCR buffer
  7. Sodium acetate
  8. dNTPs
  9. Agarose gel
  10. TE-saturated phenol
  11. Ethidium bromide
  12. DDW
  13. Stock solution A (see Recipes)
  14. Master mixture A (see Recipes)

Equipment

  1. 1.5 ml Eppendorf tube
  2. Centrifuges
  3. Micropipette

Procedure

Copyright: © 2012 The Authors; exclusive licensee Bio-protocol LLC.

Category

Molecular Biology > DNA > PCR

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