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Hence, the objective of this study was to develop a sensitive and reliable PCR-based method for detection of C. sakazakii by screening for specific target genes.
All reactions were two-step beginning with cDNA synthesis followed by conventional or quantitative PCR for specific target genes.
(2) Genome navigation uses the GA-ribbon as a search path for specific target genes that is up to 730-fold shorter than the full-length chromosome.
PCR for specific target genes was performed using the following thermal cycling conditions, 95°C -7' initial activation followed by 94°C-30 94°C-300";72°C-45", for 357°C-30 72°C-45the Immomix PCR mastermix from Bioline (Randolph, MA).
Primer pairs for specific target genes were designed as listed in Additional file 1: Table S1.
This result illustrate that designing qPCR primers close to the 3' end is critical for specific target genes.
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Subsets of RNAs were used for determining transcript levels of specific target genes by RT-qPCR.
UPF1 is known to be involved in RNA degradation pathways for some specific target genes, but it could also be a universal stabilizing protein.
Analyses of the target genes specific for either COPD or lung cancer indicated that ITGA5 was as the only specific target for the COPD, whereas 14 specific target genes were found for lung cancer with 16 target genes shared by both diseases.
The respective analysis results indicate that we can derive specific target genes for the different diseases that may be of relevance for molecular pathological mechanisms.
In order to identify specific target genes for each cell type specific enhancer, the overlapped genes were excluded for further analysis.
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