The organization keeps on the procedure concept “scientific management, high quality and efficiency primacy, purchaser supreme for China Manufacturer for 2× Concentrated Premix for Unpurified Sample Real-Time Quantitative PCR Direct Multiplex Probe Qpcr Mix Plus U, Our business is dedicated to giving customers with significant and secure top quality products at aggressive price, making each and every customer pleased with our services.
The organization keeps on the procedure concept “scientific management, high quality and efficiency primacy, purchaser supreme for China Taq DNA Polymerase and Qpcr, Our company has abundant strength and possesses a steady and perfect sales network system. We wish we could establish sound business relationships with all customers from at home and abroad on the basis of mutual benefits.
Real Time PCR primer design principles

Forward Primer and Reverse Primer

For Real Time PCR, primer design is very important. Primers are related to the specificity and efficiency of PCR amplification, and can be designed with reference to the following principles:

• Primer length: 18-30bp.
•  GC content: 40-60%.
• Tm value: Primer design software, such as Primer 5, can give the Tm value of the primer. The Tm values of the upstream and downstream primers should be as close as possible. The Tm calculation formula can also be used: Tm = 4 °C (G + C) + 2 °C (A + T). When performing PCR, a temperature below the primer Tm value of 5 °C is generally selected as the annealing temperature (the corresponding increase in the annealing temperature can increase the specificity of the PCR reaction).
• Primers and PCR products:

1.  Design primer PCR amplification product length is preferably 100-150bp.
2.  Design primers in the secondary structural area of the template should be avoided as much as possible.
3. Avoid the formation of 2 or more complementary bases between the 3′ ends of upstream and downstream primers.
4.  Primer 3′ terminal base can not be present with 3 additional consecutive G or C.
5. The primers themselves cannot have complementary structures, otherwise a hairpin structure will be formed, affecting PCR amplification.
6. ATCG should be distributed as evenly as possible in the primer sequence, and the 3′ terminal base should be avoided as T. 

Appendix 1：Cell Direct RT-qPCR Kit component supplement pack

1.Cell Lysis Solution

 The organization keeps on the procedure concept “scientific management, high quality and efficiency primacy, purchaser supreme for China Manufacturer for 2× Concentrated Premix for Unpurified Sample Real-Time Quantitative PCR Direct Multiplex Probe Qpcr Mix Plus U, Our business is dedicated to giving customers with significant and secure top quality products at aggressive price, making each and every customer pleased with our services.
China Manufacturer for China Taq DNA Polymerase and Qpcr, Our company has abundant strength and possesses a steady and perfect sales network system. We wish we could establish sound business relationships with all customers from at home and abroad on the basis of mutual benefits.

Real Time PCR primer design principles

Forward Primer and Reverse Primer

For Real Time PCR, primer design is very important. Primers are related to the specificity and efficiency of PCR amplification, and can be designed with reference to the following principles:

• Primer length: 18-30bp.
•  GC content: 40-60%.
• Tm value: Primer design software, such as Primer 5, can give the Tm value of the primer. The Tm values of the upstream and downstream primers should be as close as possible. The Tm calculation formula can also be used: Tm = 4 °C (G + C) + 2 °C (A + T). When performing PCR, a temperature below the primer Tm value of 5 °C is generally selected as the annealing temperature (the corresponding increase in the annealing temperature can increase the specificity of the PCR reaction).
• Primers and PCR products:

1.  Design primer PCR amplification product length is preferably 100-150bp.
2.  Design primers in the secondary structural area of the template should be avoided as much as possible.
3. Avoid the formation of 2 or more complementary bases between the 3′ ends of upstream and downstream primers.
4.  Primer 3′ terminal base can not be present with 3 additional consecutive G or C.
5. The primers themselves cannot have complementary structures, otherwise a hairpin structure will be formed, affecting PCR amplification.
6. ATCG should be distributed as evenly as possible in the primer sequence, and the 3′ terminal base should be avoided as T. 

Appendix 1：Cell Direct RT-qPCR Kit component supplement pack

1.Cell Lysis Solution

Instruction Manuals:

 Quick Easy Cell Direct RT-qPCR Kit-Taqman