PCR (polymerase chain reaction) is a widely used technique to amplify/replicate DNA. The DNA that is replicated by the DNA polymerase is itself used for the next template for replication. This is the chain reaction that allows the initial DNA template (a single or few copies of a piece of DNA) to be exponentially amplified. Standard PCR uses methods to detect PCR amplification at the endpoint of the PCR reaction.
qPCR, also called real-time PCR, incorporates either dyes or probes to allow for the detection of PCR amplification during the actual reaction. More sensitive than standard PCR, qPCR measures the exponential phase of PCR for more accurate quantitation.
RT-PCR (reverse transcription – polymerase chain reaction) uses mRNA as the input sample. A reverse transcriptase enzyme is used to generate cDNA from the RNA template. Subsequently, the cDNA is used as a template for either PCR or qPCR.
Our AMPIGENE® Taq Mixes and DNA Polymerases all include stabilizers, enhancers, buffers, polymerases, dNTPs, and magnesium to enable easy standard PCR. The AMPIGENE® HS Taq Mixes and HS DNA Polymerase allow for more sample types and increase PCR sensitivity.
AMPIGENE® 1-Step RT-PCR Kit
Convenient and easy, our AMPIGENE® 1-Step RT-PCR Kit has all the components for efficient cDNA synthesis and PCR in one tube. The MMLV RTase is thermostable and extremely active, allowing efficient cDNA synthesis, while the buffer contains enhancers to prevent the formation of primers/dimers to increase sensitivity and specificity.
AMPIGENE® qPCR Mixes
Using the latest developments in polymerase technology and buffer chemistry, AMPIGENE® qPCR Green Mixes and AMPIGENE® qPCR Probe Mixes require minimal optimization to deliver high performing real-time PCR. For ease of use, the qPCR Mixes come premixed with a range of different ROX reference dye intensities in order to suit your qPCR instrumentation.
AMPIGENE® qPCR 1-Step Kits
Our AMPIGENE® qPCR 1-Step Kits contain components needed for convenient cDNA synthesis and qPCR in one tube. The MMLV RTase is optimized for efficient cDNA synthesis while the qPCR components allow sensitive quantitation. Choose the convenient AMPIGENE® qPCR 1-Step Kit in the optimized reference dye format for your qPCR needs.
Higher Yield
450bp fragment of the human myc gene was amplified with AmpiGene® Taq DNA Polymerase and compared with Taq Polymerases from other suppliers. A serial dilution of human genomic DNA was incubated under standard PCR conditions. AmpiGene® Taq delivers higher yield and sensitivity as compared with all six competing products.
Higher Sensitivity
Human Genomic DNA 100ng/µl, then 1/3 serial dilution A) β2MG (Beta-2 microglubulin) gene B) pgk gene.
AmpiGene® HS Taq DNA Polymerase does not produce primer-dimers and can amplify lower concentrations of DNA even from genes such as pgk that have high secondary structure levels.
Instrument Compatibility of AMPIGENE® qPCR Products
AMPIGENE® qPCR Probe Mixes and 1-Step Kits are available in Lo-ROX, HI-ROX, and No-ROX.
AMPIGENE® qPCR Green Mixes and 1-Step Kits are available in Lo-ROX and HI-ROX.
Nucleic acid dye for gel staining and higher sensitivity qPCR
CYGREEN® Nucleic Acid Dye is a DNA intercalating agent that is used to stain DNA. The DNA-dye complex emits a fluorescence spectra that makes it suitable for qPCR and gel staining applications. CYGREEN® Nucleic Acid Dye has excitation/emission spectra similar to commonly used qPCR dyes and can be substituted in to increase sensitivity and reduce cost.
Higher sensitivity allows detection of low target concentration
Greater PCR efficiency for accurate and reproducible results
Soluble in water for higher stability
Compatible with various enzyme systems
Suitable for commonly used rotor-type and plate-type qPCR platforms
Validated for qPCR applications and gel staining
Lower cost per assay compared to commonly used dyes
CYGREEN® Nucleic Acid Dye has higher sensitivity than competitor dyes.
CYGREEN® Nucleic Acid Dye and 4 other competitor dyes were used in qPCR reactions with varying target concentrations.
EQ Quencher Dyes for FRET Applications
EQ quencher dyes are suitable as acceptors in fluorescence resonance energy transfer (FRET) applications due to their broad visible absorption but no detectable fluorescence emission.
Normalized absorbance of EQ dyes showing their range of quenching through FRET.
DNA Decontamination Solutions
The PCR Decontamination Kit removes DNA contamination from PCR mastermixes. Contaminating DNA can be removed in the presence of primers and probe by utilizing a specific dsDNase. The dsDNase is irreversibly inactivated by heating, leaving the mastermix components unaffected.
Reduce background to improve target detection
No effect on PCR sensitivity
Easy protocol – decontaminate directly in mastermix
The gDNA Removal Kit removes DNA contamination from RNA prior to reverse transcription. It uses a heat labile dsDNase which is irreversibly inactivated at low temperatures. This enables an inactivation step which is gentle enough to preserve both quality and quantity of all present RNA.
Removes gDNA from RNA prior to reverse transcription in the same tube
HL-dsDNase is inactivated without reducing quality or quantity of RNA
Minimizes pipetting steps and suitable for high throughput experiments
Efficient removal of gDNA without affecting RNA
The gDNA Removal Kit removes gDNA from RNA preps to levels below the detection limit but does not affect RNA. Compared to untreated control, no reduction in cDNA level (SDHA) was observed RT-qPCR.
PCR Clean-up
PCR & Gel clean-up columns use silica membrane technology in a mini spin column format to deliver easy and efficient DNA purification from PCR products, agarose gels, and labeling reactions.
High recoveries for fragments down to 50 bp
Small elution volume down to 15 µl
Binding buffer with pH indicator
Shrimp Alkaline Phosphatase, (recombinant), is a heat labile alkaline phosphatase that can be used in a PCR clean-up application to dephosphorylate nucleotides. It is fully inactivated by a short heat treatment to simplify workflows.
Completely heat inactivated after 5 min at 65°C
Highly stable at room temperature and 4°C
Rapid and complete heat inactivation
Heat inactivation of Shrimp Alkaline Phosphatase at 65°C and 75°C