qRT-PCR Direct from Cells













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Nucleic Acid Amplification & Expression Profiling

CellsDirect™ One-Step and Two-Step qRT-PCR Kits

Sensitive qPCR results, faster throughput with limited samples

CellsDirect™ qRT-PCR kits deliver highly sensitive and specific, real-time qPCR results directly from cells, without the need for an RNA purification step when testing with less than 10,000 cells per reaction to as low as 1 cell. CellsDirect™ technology is designed for maximum sensitivity with small samples (Figure 1). By eliminating costly, time-consuming RNA purification procedures (Figure 2), it is especially valuable for high-throughput applications and is ideal for gene expression and RNAi knockdown experiments.

  • Increased sensitivity - No RNA isolation steps to avoid sample loss and PCR inhibition - use of SSIII for extensive sensitivity down to 1 cell / reaction.
  • Significant savings - Less hands-on time and a lower cost than RNA purification kits
  • Robust qPCR results - Real-time detection of targets from a single cell to 10,000 cells
  • Versatility - Compatible with a wide variety of cell lines and samples from Laser Capture Microdissection (LCM)
CellDirect Detection
Figure 1. CellsDirect™ qRT-PCR kits provide sensitive detection down to a single cell

HeLa cells were lysed and resuspended according to the CellsDirect™ protocol. Ten-fold serial dilutions of cell lysate were prepared from 10,000 cells to one cell. A low abundance target, RRKCA, was detected using a TaqMan® probe with the CellsDirect™ One-Step qRT-PCR Kit with ROX on an ABI PRISM® 7700 instrument. Efficiency as measured from a standard curve (not shown) was 97%, with an R² value of 0.998.

CellsDirect Throughput
Figure 2. CellsDirect™ technology increases throughput

CellsDirect™ qRT-PCR technology provides a significant savings of hands-on time and cost compared to conventional qRT-PCR methods involving expensive and lengthy RNA purification
steps.

CellsDirect™ One-Step qRT-PCR Kits

CellsDirect™ One-Step qRT-PCR kits combine sensitivity with the highest level of convenience and throughput. With CellsDirect™ One-Step Kits, just add your cell lysate directly to your qRT-PCR reaction mixture, place it in your real-time instrument, and go. These kits are ideal for gene ex­pression assays from samples collected using LCM (Figure 3).

CellsDirect Sensitive Result
Figure 3. CellsDirect™ One-Step qRT-PCR Kits provide sensitive results with LCM samples

CellsDirect™ Two-Step qRT-PCR Kits

CellsDirect™ Two-Step qRT-PCR Kits provide the sensitivity of CellsDirect™ technology (Figure 4) while allowing archiving of cDNA. Two-step kits are available for either probe/primer or SYBR® Green detection.

CellsDirect Two Step
Figure 4. CellsDirect™ Two-Step qRT-PCR Kit detects from one to 10,000 cells