500 ng of cDNA was used as a template for performing qRT-PCR using SYBR Green Supermix (Bio-Rad, CA, USA) on ABI Prism 7500 sequence detection system (Applied Biosystems, CA, USA).
500 ng of cDNA was used as a template for performing qRT-PCR using SYBR Green Supermix (Bio-Rad, Hercules, CA) on ABI Prism 7700 sequence detection system (Applied Biosystems, Foster City, CA, USA).
The FTA Micro Card was excluded after the first phase of experiments due to the limitation of paper size, which provided insufficient DNA template for performing the necessary number of single PCR assays required as a diagnostic routine for bacterial meningitis in a clinical setting.
pTNFSF9 (3) was used as a template for mutagenesis performed by the QuickChange site-directed mutagenesis method (Stratagene).
The resulting first-strand cDNA (0.5 μl of the reverse transcription reaction) was used as a template for PCR, performed with Taq polymerase (Promega, Madison, WI, USA) and specific primers for Arx, Crh, Ghrh, Gsh1, Sim1, and Trh mRNAs.
The resulting first-strand cDNA (0.5 μL of the reverse transcription reaction) was used as a template for PCR, performed with Taq polymerase (M8305, Promega, Madison, WI) and specific primers for Gata2 (forward primer: 5′-cttcctccagtctctcttttgg-3′, reverse primer: 5′-tacaccagctttggcctctg-3′) and Gata3 (forward primer: 5′-ctgcaaaccattaaacga-3′, reverse primer: 5′-acgtctccagcttcatgctatc) mRNAs.
In short, a realisation that a one-size-fits-all template for getting poorly performing schools into an adequate state doesn't then provide the broader, stretching education we all want to see.
During the past six decades, the Toyota Motor Company established a 'lean' production and management system (the Toyota Way), which has become an iconic template for a high performing and learning organization.
PCR for amplifying total cDNA as the template was performed for up to 30 cycles with denaturation for 30 s at 95°C, annealing for 45 s at 64°C and elongation for 60 s at 72°C with the Advantage™ 2 PCR Enzyme Systems (Takara, Otsu, Japan).
To remove this effect, first, we mixed all reads that were sorted to two indices for each template, and performed clustering for these mixed reads.
We then enriched for cDNA templates by performing multiplex incorporating PCR reactions (≤18 cycles), and isolating 250 550 base pair fragments by gel purification.
