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About Sequence Detection Chemistries

 Overview Applied Biosystems has developed two types of chemistries used to detect PCR products using Sequence Detection Systems (SDS) instruments:

  • TaqMan ® chemistry (also known as “fluorogenic 5´ nuclease chemistry”)
  • SYBR ® Green I dye chemistry

 TaqMan® Chemistry

The TaqMan chemistry uses a fluorogenic probe to enable the detection of a specific PCR product as it accumulates during PCR cycles.

 Assay Types that Use TaqMan Chemistry

The TaqMan chemistry can be used for the following assay types:

  • Quantitation, including:
    • One-step RT-PCR for RNA quantitation
    • Two-step RT-PCR for RNA quantitation
    • DNA/cDNA quantitation
  • Allelic Discrimination
  • Plus/Minus using an IPC

SYBR Green I Dye Chemistry

The SYBR Green I dye chemistry uses SYBR Green I dye, a highly specific, double-stranded DNA binding dye, to detect PCR product as it accumulates during PCR cycles.

The most important difference between the TaqMan and SYBR Green I dye chemistries is that the SYBR Green I dye chemistry will detect all double-stranded DNA, including non-specific reaction products. A well-optimized reaction is therefore essential for accurate results.

 Assay Types that Use SYBR Green I Dye Chemistry

The SYBR Green I dye chemistry can be used for the following assay types:

  • • Quantitation, including:
    • One-step RT-PCR for RNA quantitation
    • Two-step RT-PCR for RNA quantitation
    • DNA/cDNA quantitation

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