RT² lncRNA PCR Array Modification

RT² lncRNA PCR Arrays are panels of RT² lncRNA qPCR Assays for long noncoding RNAs related by pathway or disease. Assays are designed using a computer algorithm, which has been developed using an in vitro assay to ensure that the resulting primer sequences generate a single PCR product of the predicted size without primer-dimers or off-target amplification – in 33 cycles of PCR amplification. The assay also ensures that the amplification efficiency of the primers is 100 +/– 10%. As a result, the algorithm designs highly effective primer sequences for SYBR^® Green based real-time PCR detection.

For optimal performance, RT² lncRNA PCR Arrays should be used together with the RT² First Strand Kit for cDNA synthesis and RT² SYBR Green Mastermixes for qPCR. These reagents have been formulated and pretested together with RT² lncRNA PCR Arrays. The RT² First Strand Kit includes a proprietary genomic DNA elimination step to remove any residual contamination in RNA samples before reverse transcription, thereby eliminating false positive signals. In addition, an artificial RNA control template is also included which can be used to follow the reverse transcription efficiency across samples and monitor against RNase contamination. Each of the real-time instrument-specific RT² SYBR Green Mastermixes contains SYBR Green and an appropriate reference dye to match the instrumentation available in your laboratory. RT² SYBR Green Mastermixes are available for all real-time PCR instruments from QIAGEN, Applied Biosystems, Bio-Rad, Stratagene, Eppendorf, Roche and other major suppliers.

Simply mix the cDNA template with the appropriate ready-to-use PCR mastermix, aliquot equal volumes to each well of the same plate and run the real-time PCR cycling program (see flowchart "Simple procedure"). RT² lncRNA PCR Arrays are compatible with all QIAGEN, ABI, Bio-Rad, Eppendorf, Roche and Stratagene instruments.

Flexible layout and controls
RT² lncRNA PCR Array Modification is available in 96-well plate, 384-well plate, and 100-well disc formats, and is used to monitor the expression of 84 lncRNA genes related to a disease state or pathway, which includes up to 4 lncRNA genes of the user's choosing that will replace assays on a cataloged array(see "Modified PCR array 96-well plate layout").
Five reference genes are included, as well as control elements for:

• Data normalization
• Genomic DNA contamination detection
• RNA sample quality
• General PCR performance
• Easy-to-use data analysis

Easy-to-use data analysis
Data can be analyzed using an easy-to-use Excel-based data analysis template or Web-based software. Data analysis is based on the ΔΔC_T method with normalization of the raw data to either housekeeping genes.

RT² lncRNA PCR Array Modification is highly suited for lncRNA research that requires the study of a few specific lncRNAs in combination with overall pathway or disease profiling.