Application:
High performance PCR reagent for probe based quantitative PCR including DNA quantification, 2-step RT PCR, SNP analysis etc. Uses only 47% of standard PCR running time. This reaction mix does not contain internal passive reference dye (ROX). It is manufactured for use with ABI real-time PCR machines.

Recommended storage conditions:
This product should be stored at –20ºC. To avoid repeated freeze-thaw, open vials should be kept at 4ºC.

Primer and probe design:
To achieve the best performance, appropriate software, such as ABI Primer ExpressTM, should be used.

1. Tm: 60ºC for primers and 68ºC~70ºC for probes with 17~30 nucleotides in length
2. Amplicon size should be small, <150bp
3. To avoid secondary structures in primers and probes
4. To avoid more than 3 consecutive Gs in primers and probes
5. Primers should not have complementary 3’ ends
6. 17 ~ 30 nucleotide in length
 

Recommended reaction conditions:
95ºC, 5min. -> (95ºC, 5sec. -> 60ºC, 30sec.) for 50 cycles.

Notes:
To achieve accurate quantification, it is highly recommended to do replicates. There is the minimal number of replicates to obtain a standard deviation. It is important to reduce pipetting error. Three ways to minimize pipetting error are:
-To prepare an amplicon specific master mix that includes CAD PCR reaction mix, primers, and probe.
-To use a repeat pipet
-To pipet volume within manufacture’s suggested range

Reference:
Mackey J, Darfler M, Nisson P, Rashtchian A.; Anal Biochem. 1993 Aug 1; 212(2): 428-35.
 

This product is distributed for laboratory research only.
Caution: Not for diagnostic use