SsoFast™ Probes Supermix

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Overview

Duplex qPCR Results 
Duplex qPCR Under Challenging Conditions
 
Gene expression analysis of two targets using SsoFast™ probes supermix on the CFX96™ real-time PCR detection system. cDNA from human liver (100 ng) was used in each 20 µl reaction. A, HEX-labeled GAPDH probe simplex reaction (■); B, Texas Red–labeled IL-2 probe simplex reaction (■); C, HEX-labeled GAPDH probe duplex reaction (■) and Texas Red–labeled IL-2 probe duplex reaction (■). Panel D illustrates an overlay of all panels to demonstrate overlapping traces and no quantification cycle (Cq) differences. No difference is observed for the detection of a low-expressing gene (IL-2) in a duplex or singleplex reaction in the presence of a high-expressing gene (GAPDH). Total qPCR run time = 38 min. RFU, relative fluorescence units.
High-Throughput qPCR 
Exceptional Stability for High-Throughput qPCR

 

SsoFast™ probes supermix maintains exceptional stability on the high-throughput CFX automation system. Tenfold serial dilutions of 100 ng to 1 pg of cDNA from human spleen were used in each 20 µl reaction to detect GAPDH. All reactions were assembled and loaded onto the CFX automation system. The following cycling conditions were used: 95°C for 10 min, followed by 35 cycles of 95°C for 15 sec and 60°C for 60 sec. After each plate run, an additional 2 hr hold time was introduced to prolong the total time between first and last plates to >48 hr. Results for five plates are shown below. RFU, relative fluorescence units.

 

Panel Plate # Start Time PCR Efficiency R2
A 1 0 hr 97.0% 0.999
B 5 14.0 hr 97.6% 0.999
C 8 24.5 hr 95.9% 0.999
D 12 39.5 hr 97.3% 0.999
E 15 48.0 hr 95.9% 0.999
Reproducibility 
Consistent Reproducibility and Quantitative Discrimination

 
SsoFast™ probes supermix demonstrates superior reproducibility. Definitive discrimination and consistent quantification can be obtained from 1.33-fold serial dilutions of input template. The GAPDH gene was amplified from varying amounts of HeLa cDNA (1 ng to 100 pg). From left to right: (■) 1 ng, 565 pg, 320 pg, 181 pg, and 102 pg; (■) 752 pg, 425 pg, 240 pg, and 136 pg. GAPDH efficiency = 91.5%, R2 = 0.997. Insert shows the standard curve for the various dilutions. Total qPCR run time = 50 min. RFU, relative fluorescence units.
Varying Cycling Conditions
Excellent Performance Under Varying Cycling Conditions

SsoFast™ probes supermix delivers robust results in a broad range of cycling conditions. Tenfold serial dilutions of 100 ng to 1 pg of cDNA from HeLa cells were used in each 20 µl reaction to detect α-tubulin. RFU, relative fluorescence units.
 
Panel Cycling Protocol qPCR Run Time
A Fast 2-step 38 min
B Standard 2-step 59 min
C Standard 3-step 67 min

SsoFast probes supermix is used in real-time quantitative PCR assays on fast cycling real-time thermal cycler platforms using fluorescent probe detection chemistry. The combination of the patented* Sso7d fusion protein technology and optimized formulation helps perform fast duplex qPCR, while maintaining a broad dynamic range, reliability, and accurate quantification for robust gene expression analysis.

  • Fast qPCR results ─ Sso7d fusion polymerase’s stringent hot-start and rapid polymerization kinetics provide fast results in less than 30 min
  • Universal conditions ─ fully compatible with your pre-existing cycling protocols and primer/probe conditions, without the need for additional optimization
  • Minimal PCR inhibition ─ high-tolerant polymerase exhibits low PCR inhibition for better performance with fast-cycling protocols
  • Distinct target detection ─ robust, simultaneous detection of up to 2 different gene targets using fluorogenic probes under fast qPCR conditions

Check out the other members of the SsoFast family that utilize the patented Sso7d fusion protein technology to deliver excellent performance in a wide range of qPCR applications ─ SsoFast EvaGreen supermix and SsoFast EvaGreen supermix with low ROX.

* U.S. patents 6,627,424 and 7,541,170.

Product expiration (after ship date) 12 months
Storage at +4°C (after thawing) Up to 6 months
Customer guarantee (minimum usage time) 12 months
SsoFast™ Probes Supermix

172-5230
2 ml, 2x real-time PCR mix, contains dNTPs, Sso7d fusion polymerase, MgCl2, stabilizers, for 200 x 20 µl reactions

List Price:   $145.00
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SsoFast Probes Supermix

172-5231
5 ml, 2x real-time PCR mix, contains dNTPs, Sso7d fusion polymerase, MgCl2, stabilizers, for 500 x 20 µl reactions

List Price:   $325.00
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SsoFast Probes Supermix

172-5232
10 ml, 2x real-time PCR mix, contains dNTPs, Sso7d fusion polymerase, MgCl2, stabilizers, for 1,000 x 20 µl reactions

List Price:   $625.00
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SsoFast Probes Supermix

172-5233
20 ml, 2x real-time PCR mix, contains dNTPs, Sso7d fusion polymerase, MgCl2, stabilizers, for 2,000 x 20 µl reactions

List Price:   $1,195.00
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Number Description Options
6252 Reagent Comparison Guide for Real-Time PCR Brochure, Rev C Click to download
6090 Amplification Reagents and Plastics Brochure, Rev D Click to download [ Add to Cart ]
10014649 Instruction Manual, SsoFast Probes Supermix, Rev B Click to download
5869 SsoFast Probes Supermix Flier, Rev B Click to download [ Add to Cart ]
5990 Gene Expression Analysis Solutions Brochure, Rev C Click to download [ Add to Cart ]