Wednesday, January 6, 2010

High dynamic range and high sensitivity qPCR and qRT-PCR

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High dynamic range and high sensitivity by primer sequestration
HotStart-IT® master mix series use a novel hot start method developed at USB called primer sequestration (Fig. 1). This novel hot start feature increases the specificity and sensitivity of SYBR-based qRT-PCR reactions by substantially reducing primer-dimer formation. With this method, the HotStart-IT® protein binds and sequesters primers at lower temperatures making them unavailable for use by Taq DNA Polymerase. Following the first heat denaturation step, the primer binding protein is inactivated and the primers are released.

Since side products are greatly avoided by primer sequestration, low background noise allows high sensitivity for fewer than 10 target copies, and therefore large linear dynamic range of 6 to 7 of magnitude (Fig. 2).

Complete and convenient
The master mix contains HotStart-IT® Taq polumerase and buffer solution that are optimum for qPCR and qRT-PCR. Passive reference dyes (ROXTM and fluorescein) are also provided in HotStart-IT® series. SYBR® Green is additionally included in the master mix for SYBR® Green specific reactions. On the other hand, extra reverse transcriptase and RNase inhibitors are provided for qRT-PCR.
You just simply add the primers and template to the master mix. You can also add other components for your applications. A brief protocol is already available in the kit for your reference.

Products of HotStart-IT® series:

  1. HotStart-IT® SYBR® Green qPCR Master Mix (2X) (Cat. No.: 75762)
  2. HotStart-IT® SYBR® Green qPCR Master Mix with UDG (2X) (Cat. No.: 75760
  3. HotStart-IT® Probe qPCR Master Mix (2X) (Cat. No.: 75766)
  4. HotStart-IT® Probe qPCR Master Mix with UDG (2X) (Cat. No.: 75764)
  5. HotStart-IT® SYBR® Green One-Step qRT-PCR Master Mix Kit (Cat. No.: 75770
  6. HotStart-IT® Probe One-Step qRT-PCR Master Mix Kit (Cat. No.: 75772)

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