Lyophilized Mastermix for RT-PCR

Description
RT-PCR is used to amplify double-stranded DNA from single-stranded RNA templates. In the RT step the reverse transcriptase synthesizes single-stranded DNA molecules (cDNA) complementary to the RNA template. In the first cycle of the PCR step Taq DNA polymerase synthesizes DNA molecules complementary to the cDNA, thus generating a double-stranded DNA template. During subsequent rounds of cycling the DNA polymerase exponentially amplifies this double-stranded DNA template.

LYO RT-PCR Mastermix is designed for performing maximum sensitive and specific RT-PCRs convenient in single tubes. The enzyme mix is based on a genetically engineered RT polymerase with enhanced thermal stability resulting in an increased specificity, higher cDNA yield and an improved efficiency for highly structured and long cDNA fragments.

The lyophilized Mastermix contains all reagents required for successful RT-PCR (except template and primer) in one tube to ensure fast and easy preparation with a minimum of pipetting steps. The premium quality enzyme mix, ultrapure dNTPs and the optimized complete reaction buffer ensure superior amplification results in simple steps. 

In LYO RT-PCR Mastermix all components of RT and PCR are mixed in one tube prior to starting the reaction and thus carried out sequentially without opening the tube. This offers tremendous convenience when applied to analysis of single targets from multiple samples of RNA and minimizes the risk of contaminations.

Handling
LYO RT-PCR Mastermix is delivered in PCR reaction tube strips or 96-well plates preloaded with a complete qPCR master mix in a dry, room temperature stable format.
The lyophilized Mixture combines highest performance with convenience of use and stability. There is no need for freezing, thawing or pipetting on ice. The few remaining pipetting steps minimize the risk of errors or contaminations.
Each vial contains all components (except primers and template) required for a 20 μl RT-PCR assay. To perform the assay, only fill up the vials with a mix of primers and RNA template.

Storage and shipping temperature: between 5 – 25 °C, minimum shelf life: 16 months

Manufactured and quality-controlled in accordance with ISO 9001:2000

LYO RT-PCR Mastermix contains:
Preloaded lyophilizates containing Reverse Transcriptase, Hot Start Polymerase, dNTPs, Reaction Buffer, MgCl2 and stabilizers, PCR-grade water

Sensitivity
Targets can generally be detected from 10 pg to 500 ng polyA RNA or 10 pg to 1 μg total RNA. Even lower amounts of RNA may be successfully amplified by using highly expressed transcripts.

RT-PCR assay preparation

Note: For optimal specificity and amplification an individual optimization of the recommended parameters may be necessary. See Datasheet!