2X HotSybr Real-time PCR Kit

For Research Use Only.

Figure: Amplification of human GAPDH gene target with 2X HotSybr Real-time PCR Kit. Amplification curves are shown for tenfold dilutions of 0.0002pM to 20pM of plasmid. Insets show the dissociation profile and the standard curve data.

Description:
This is a high performance real-time PCR reagent. It utilizes MCLAB's proprietary quantitative PCR technology.

Advantages:
1. 2x HotSybr PCR Reaction Mix products cut the total reaction time down to half.


- Normally the total reaction time is 4350 seconds: 95°C, 10 minutes => (95°C, 15 seconds => 60°C, 60 seconds) x 50
- For MCLAB's 2x HotSybr PCR Reaction Mix, the total reaction time is reduced to 2350 seconds: 95°C, 10 minutes => (95°C, 5 seconds => 60°C, 30 seconds) x 50

2. One-step RT-PCR products are faster.


3. 2x HoTaq PCR Reaction Mix products are superior in amplifying difficult templates comparing with similar products from other suppliers.

- This is the amplification of GPIIB gene (70% G+C) .
- 10 ~ 10K copies from 30pg human genomic DNA have been detected.

Application:
Probe based quantitative PCR: including DNA quantification, 2-step RT PCR, SNP analysis, etc.

Primer and probe design:
Appropriate software, such as ABI Primer Express^TM, should be used.

Recommended Reaction Conditions:
95°C, 10 minutes -> (95°C, 5 seconds. -> 60°C, 30 seconds.) for 50 cycles -> melting curve.

Recommended Storage Conditions: -20°C

Notes:
To achieve accurate quantification, it is highly recommended to (1) do replicates; (2) reduce pipetting error; (3) primer concentration from 100nM to 300nM; (4) run melting curve following amplification cycles.

Reference:
Higuchi R, Dollinger G, Walsh PS, Griffith R; Bio/Technology 10: 413-417, 1992