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PCR Amplification

GC-rich Targets addressed using CleanAmpTM 7-deaza-dGTP; towards improved Genetic Disease Diagnosis.

CleanAmpTM 7-deaza-dGTP is not a name that trips easily off the tongue - that is for sure! However, this addition to TriLink Biotechnologies's range of custom nucleic acid-based products has an importance that transcends its description as a ‘Hot Start solution for the Polymerase Chain Reaction (PCR) amplification of problematic GC-rich sequences'. CleanAmpTM 7-deaza-dGTP is significant to one of the deadliest of diseases still troubling mankind: tuberculosis. 

Tuberculosis is not just a disease of the 19th century - the province of romantic poets and corseted young women - it is a disease that is estimated to infect one third of the world's population, including, increasingly, populations in what is generally referred to as the developed world.

Leprosy is another disease of pathogenic mycobacteria that - like tuberculosis - causes misery and death over large swathes of the world. Anything that can aid the rapid diagnosis of these scourges is a great addition to our toolbox of medical techniques that help combat suffering. Disease diagnosis using the techniques of genetics is a rapidly advancing area with enormous potential to address serious and costly, both in human and economic terms, diseases.

PCR amplification, specifically Hot Start PCR, whereby even single strands of DNA can be efficiently multiplied across several orders of magnitude, is a genetic technique that should be enormously effective for diagnostic purposes. There are complications though. The bacterial cell's chromosome unlike a ‘normal' cell is not found in a defined nucleus - although given their environmental prevalence the bacterial chromosome might be considered the normal case! Critically, in the case of mycobacteria, their DNA also contains a higher proportion of the guanine-cytosine (GC) base pairing than a ‘normal' cell.

This is pertinent to PCR because DNA targets that are rich in GC sequences are problematic - initially there can be inadequate DNA strand separation and complex secondary structures can be formed during the process.  

CleanAmpTM 7-deaza-dGTP is a combination of two techniques designed to improve GC-rich target amplification. CleanAmpTM, TriLinks Hot Start approach to PCR, is a method of limiting active reagent concentrations during the early cycles of amplification, when the target is in low concentration, thus reducing background amplification. The nucleotide analog 7-deaza-dGTP helps reduce the occurrence of secondary structures during amplification.

Together, especially in the CleanAmpTM 7-deaza-dGTP Mix, for especially challenging targets with significantly higher GC-content, they represent a important contribution to the genetic diagnosis of disease.

CleanAmpTM 7-deaza-dGTP, not easy to say, but well worth remembering.

Date added: 2011-07-13 11:52:27