DESCRIPTION

Realtime PCR Master Mix series is a Taq DNA polymerase-based 2 x master mix for real-time PCR, which contains all components, except for the primer and probe. Realtime PCR Master Mix is applicable in TaqMan™ assays or hybridization probe assays, in combination with each probe. SYBR Green™ Realtime PCR Master Mix is applicable for intercalation assay with SYBR® Green I. SYBR Green™ Realtime PCR Master Mix -Plus- is effective for reducing non-specific amplifications or primer dimers in the intercalation assay.

FEATURES

  • Can be used in glass capillary systems (e.g. LightCycler, Roche Molecular Systems, Inc.).
  • Can be used in a passive reference system (e.g., ABI PRISM™, Applied Biosystems, Inc.). The passive reference dye does not affect any other systems.
  • Hot Start technology with anti-Taq DNA polymerase antibodies enables high specificity and reproducible amplification.
  • ffective for reducing non-specific amplifications or primer dimers. <<SYBR Green™ Realtime PCR Master Mix -Plus->>

APPLICATIONS

<Realtime PCR Master Mix>
TaqMan™ assays or hybridization probe assays

<SYBR® Green Realtime PCR Master Mix>
Intercalation assay with SYBR® Green I

<SYBR® Green Realtime PCR Master Mix -Plus->
Intercalation assay with SYBR® Green I with high specificity

STORAGE CONDITION

Store at -20ºC

COMPONENTS

The reagent includes the following components for 200 reactions (QPK-101,QPK-201, QPK-212), 50 µL total reaction volume:

<QPK-101>
Realtime PCR Master Mix 1 mL x 5
<QPK-201>
SYBR® Green Realtime PCR Master Mix 1 mL x 5
<QPK-212>
SYBR® Green Realtime PCR Master Mix -Plus- 1 mL x 5
Plus solution 1 mL

APPLICATION DATA

Example 1. Comparison of the sensitivity and efficiency of the SYBR® Green assay

Amplification of the β-actin gene was detected using serially diluted genomic DNA solutions (10n dilution; 30 ng- 3 µg) with real-time PCR kits for the SYBR® Green assay. As shown in Fig. 1, SYBR® Green Realtime PCR Master Mix [Code No. QPK-201] showed greater sensitivity and efficiency than other kits (companies A and B). Moreover, SYBR® Green Realtime PCR Master Mix [Code No. QPK-201] generated fewer primer dimers than the other kits.

Fig. 1 Comparison of the amplification and melting curves

Example 2. TaqMan™ assay using Realtime PCR Master Mix

Amplification of β-actin mRNA was detected using serially diluted total RNA (5n dilution; 6.4 ng-100 µg) with TaqMan™ probe. As shown in Fig. 2, clear amplification curves were obtained.

Fig. 2 Amplification curve (ABI PRISM™ 7700)

Example 3. Comparison of specific activity

Amplification of an 80-bp target gene was detected using serially diluted rodent genomic DNA (10n dilution; 0.01 ng-10 ng) with SYBR® Green Realtime PCR kits. As shown in Fig. 3, SYBR® Green Realtime PCR Master Mix Plus [Code No. QPK-212] showed greater specificity and dynamic range than the other kit (company A).

Fig. 3 Comparison of the amplification and melting curves