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Universal PCR primer pair GAF (F): 5'-GAG GAA GTA AAA GTC GTT AAC AAG GTT TG-3' and GAF (R): 5'-GAA ATT CAC AAA GGG TAG GAT GAT TT-3' was used to amplify a specific region of 18S rDNA from rumen anaerobic fungi.
Reactions were performed in 20 [micro]L of 1xTaqMan Universal PCR Master Mix (Applied Biosystems, Foster City, CA, USA), 900 nmol/L of LT.
However, the relative risk of contamination remains the major problem of using universal PCR alone as a specific test to confirm a diagnosis (9).
The TaqMan[R] Universal PCR MasterMix (ABI 4324018) containing AmpliTaq Gold DNA Polymerase, AmpErase without UNG, dNTPs with dUTP, and buffer components optimized as a 2x solution.
A characterization and evaluation report for Neo-EpCAM(TM) has been completed in universal PCR and ELISA.
Discussion Due to the problems associated with the development of a sensitive and specific universal PCR assay (10), a single-tube multiplex PCR was developed based on N.
Amplification reactions were performed by using 5 [micro]L of extracted nucleic acids with the TaqMan Fast Universal PCR kit (Applied Biosystems) in a final volume of 20 [micro]L.
To further improve the reproducibility of PCR, manufacturers have introduced universal PCR master mixes containing all reagents at predetermined concentrations.