Real-time-PCR was performed in a reaction volume of 20 μl containing 2 μl cDNA, Light Cycler TaqMan Master (Roche), primers and probes for p21, EGFR and the housekeeping gene porphobilinogen deaminase (PBGD) in concentrations recommended by the manufacturer (Real Time Ready Assays, Roche...
Briefly, the DNA was mixed with the primer pool, containing all primers for generating the 224 amplicons and the AmpliSeq HiFi Master Mix, and transferred to a PCR cycler (BioRad, Munich, Germany). PCR cycling conditions were as follows: Initial denaturation: 99°C for 2 min, cycling: ...
0.5 to 2 μg mRNA was reverse transcribed in a reaction mix containing 10U/μL Moloney Murine Leukemia Virus reverse transcriptase (Life Technologies), 5 ng/ μL Random Primers (Life Technologies), 1x first strand buffer (Life Technologies), 5 mM DTT (Life Technologies),...
Se inicio cribado de amiloidosis cardiaca según práctica clínica habitual de cada centro; las gammagrafías positivas se analizaron centralmente. Resultados Se incluyeron 422 pacientes, 387 de los cuales evaluados para amiloidosis cardiaca. Un total de 65 pacientes (16,8%) se diagnostic...