RT-PCR kits

Real-time quantitative PCR
Real-time quantitative PCR (RT-qPCR) is an innovative and reliable technique for quantitative analysis of gene expression, mutation detection, allele discrimination and single nucleotide polymorphisms (SNP) genotyping. Contrary to classical PCR technology, real-time quantitative PCR can not only detect the presence of a particular DNA sequence at the end of the PCR reaction but also monitor and quantify the amount of amplification in real time.
RT-qPCR systems determine the amount of amplified product by measuring the amount of light emitted by a fluorescent dye. The emission intensity is in direct proportion to the amount of amplified PCR product and, therefore, quantifies the amount of target DNA. The whole RT-qPCR assay can be directly performed in a sealed tube without any purification or separation steps, avoiding risk of contamination and post-PCR handling.
Because double-stranded DNA products have their own specific melting temperature, RT-qPCR systems also enable mutation detection via melting-curve analysis. Furthermore, analysis of the melting-curve data enables one to distinguish between specific PCR products and non-specific PCR products.

Sequence-specific Probes and Non-Sequence-Specific Probes
Detection by RT-qPCR can be sequence-specific or non-sequence specific, and the method chosen can have an influence on your outcome. For instance, SYBR® Green 1, a non-sequence-specific reagent, can bind to any double-stranded DNA including primer-dimers and other non-specific reaction products, and so may lead to an over-estimation of the amount of target DNA. However, sequence-specific probes such as TaqMan or Molecular beacon probes (both types of dual-labeled fluorescent probes) bind only to the target template and therefore achieve accurate and reliable quantification of the amount of target DNA.

Design and Synthesis Service of RT-PCR Primers 
GenePharma provides free design of RT-PRC primers and probes. 
Our optimized primer design techniques - utilizing the latest database resources and bioinformatics knowledge - enable us to create and select the best primers. You may get the best experimental results by using primers and RT-PCR reagents provided by our company in our recommended standard reaction conditions. Our special primer design has the following characteristics:

1. Primer-dimer formation is very unlikely to occur;
2. Primers are normally designed crossing exon junction, which makes amplification of genomic DNA very unlikely;
3. The primer sequence does not contain known SNP sites (confirmed by dbSNP);
4. The specificity is confirmed through homology search;
5. Choosing the design region under experimental purposes;
6. Normally, 1 - 3 pairs of corresponding primers are provided for one target gene;
7. Design according to the full sequence of target gene;
8. Design among the 1,500 bases at the 5’ end;
9. Design among the 1,500 bases at the 3’ end.

Customers must only provide the following information: 

1. Species (such as Human, Mouse, Rat);
2. Target gene (GeneBank Acc, GeneID, key word);
3. Primer design region (full sequence, 5 ' end sequence, 3 ' end sequence);
4. Non-sequence-specific (SYBR Green I) or sequence-specific fluorescent probe (TaqMan probes or Molecular beacons, see the table below for our available probe types)

GenePharma available RT-qPCR fluorescent probe types :

For technical assistance, email support@genepharma.com. For questions related to order placement, email bd@genepharma.com.