Can I use the original Quant-iT™ Kits with the Qubit® Fluorometer?

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No, we do not recommend this. Some of the dyes in the original Quant-iT™ kits (those NOT listed as “for use with the Qubit® fluorometer”) are not compatible with the Qubit® Fluorometer. In addition, the new Quant-iT™ kits (labeled as “for use with the Qubit® Fluorometer”) have standards formulated to be more accurate with the Qubit® fluorometer. The Qubit® Fluorometer-compatible kits are also less expensive per assay if you are processing fewer than 20 samples at a time.

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Can I use the Quant-iT™ and Qubit® Kits with other fluorometers?

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All Quant-iT™ and Qubit® kits are compatible with all fluorometers and microplate readers that have the appropriate light sources and filters. You won’t have access to the mathematical algorithm in the Qubit® kit for generating the most accurate standard curve, so you just want to make a few dilutions of the highest standard curve in the kit so you can generate a curve from more points. This will give you better accuracy on another fluorometer.

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Can the Qubit® kit give an indication of sample quality?

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Yes, but the Qubit® kit does not give an A260/A280 ratio. The A260/A280 ratio is rather a vague measurement and there is no way to determine what the ratio 1.7 really means. However with the Quant-iT™ dyes, you will be able to determine a true ratio of DNA to RNA, find out how much DNA or RNA you have, and how much contamination nucleic acid you have. You can compare the Quant-iT™ dye reading to your A260 reading. If the A260 reading is higher than the concentration obtained from the Quant-iT™ assay (which often happens), then you have contamination.

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Why are my UV absorbance readings higher than the Qubit® platform readings?

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UV absorbance readings will measure DNA, RNA, proteins, free nucleotides, excess salt, etc. Everything and anything that absorbs at 260 nm, in fact. Since the Qubit® Platform uses fluorescent dyes that preferentially bind DNA, RNA, or protein to generate a fluorescent signal, your concentration data will be more accurate than that obtained traditionally using absorbance-based methods. For additional information on the Qubit® and associated reagents, search for "Qubit" on the Life Technologies homepage.

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Can the Qubit® Quantitation Platform do a good job of quantifying plasmids?

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Yes. If you are doing a typical plasmid miniprep with lots of DNA (over 50 ng/μL), you should use the Quant-iT™ DNA BR assay, as this will allow you to quantitate without making an extra dilution. If, on the other hand, you are doing “plasmid rescue” or some other method that gives you small amounts of DNA, then you should use the Quant-iT™ DNA HS assay.

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How does the accuracy and sensitivity of the Qubit® Quantitation Platform compare to a microplate reader?

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The accuracy and sensitivity of the Qubit® Quantitation Platform is the same as that of a microplate reader. This was a requirement during product development. The detection limits for each Qubit® kit can be found on the corresponding product manual, which can be found by searching our website by keyword or catalog number.

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