Product Highlight
Improving GC-rich Target Amplification
We are proud to introduce the latest addition to the CleanAmp™ Product Line, CleanAmp™ 7-deaza-dGTP for GC-rich target amplification. When DNA targets high in GC content are amplified, PCR product formation can often be compromised by inadequate strand separation and the propensity for complex secondary structure formation. The inability of the DNA polymerase to negotiate through regions of strong secondary structure, such as hairpins, often results in the formation of truncated PCR products. In addition, mis-priming amplification products are prevalent when primers are high in GC content. Despite the inherent challenges, detection of GC-rich sequences is becoming increasingly important for the molecular diagnosis of inherited diseases. The use of standard 7-deaza-dGTP is a notable method for overcoming this problem. TriLink has developed CleanAmp™ 7-deaza-dGTP, an elegant fusion of the secondary structure reducing nucleotide analog 7-deaza-dGTP and TriLink’s CleanAmp™ dNTP Hot Start technology. CleanAmp™ 7-deaza-dGTP is available individually for amplification of routine GC-rich targets or as the CleanAmp™ 7-deaza-dGTP Mix which is recommended for more challenging targets with higher GC content.
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Research Update
Better Multiplex with CleanAmp™ Primers
Recent data demonstrates CleanAmp™ Primers’ utility in problematic multiplex systems. Like CleanAmp™ dNTPs, CleanAmp™ Primers will improve yield and specificity in multiplex systems. However, CleanAmp™ Primers offer higher specificity with a greater number of targets. In the systems shown in the figure below, CleanAmp™ Turbo Primers greatly improve formation of the desired products in multiplex PCR for seven and nine targets from genomic DNA sources. The development of a robust multiplex PCR assay typically requires an iterative design process to discover primer pairs that are both specific for the targets of interest and exhibit a low level of off-target amplicon formation. With CleanAmp™ Primers minimal design optimization is required.
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Ask An Expert Featured FAQ
Q. How do I select the CleanAmp™ Primer type that I need?
A.
There are two forms, which differ in the rate of release of the protecting group. CleanAmp™ Turbo Primers are for fast cycling (<45 minutes) and multiplexed PCR. You will see improved amplicon yield and reduced mis-priming and primer dimer formation. CleanAmp™ Precision Primers are for standard cycling (>45 minutes) and for one-step reverse-transcription PCR. You will see improved specificity of amplification, improved limit of detection and the greatest reduction in mis-priming and primer dimer formation.
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Customer Testimonial
"Our work, in part, involves development of high sensitivity tests for mutations present in only a small fraction of cells under test. This requires high specificity also and it has proven to be the case that the TriLink CleanAmp™ Primers provided the specificity needed to capitalize on qPCR's inherent high sensitivity. Using your primers we can routinely identify the presence of mutation fractions as low as 0.001%."
- John Owen M.D., Professor of Hematology & Oncology at Wake Forest University |
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