I have *what* in my sample?

Posted by John on 4 May 2015 | 1 Comments

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Reagent contamination in PCR has long been documented - not only by operators but also from commercial sources long before those operators ever laid DNA-ridden hands on them.

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The dreaded 'C'

Posted by John on 18 August 2010 | 0 Comments

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Well, its probably the same as tempting fate when mentioning 'the Scottish play' . . .but then we're not overly theatrical so we can say MacBeth. However superstition may reign when you get the band on the additional lane on the gel or the extra qPCR amplification curve and think 'did I load template in that reaction?'. That's before the repeat reactions tell you that it's con . . .contam. . .the dreaded 'c'.
Thankfully instances of it are usually rare in real-time qPCR (hopefully only the one gel to analyse products when establishing an assay) but it can be a major hassle to eliminate.
One of the difficulties can be the research desire to know where the contamination is coming from versus immediately getting back to strong positives and clean negatives. In the former, often one reagent may be exchanged at a time - often in the vain attempt to avoid throwing out enzyme vials or master mix, while spreading the contamination further.

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