Methods for Quantation of Very Low (<5pg/uL) DNA

[winterc16]

Does anyone have experience with quantifying very low input DNA amounts, specifically amounts that are less than 5pg/ul?

Currently we still prepare libraries on zero quant. amounts with varying amounts of success. This means that in some cases our initial quant. was actually zero and we just wasted a bunch of time sequencing kit. In other cases our sample did sequence even though it had a quant. of zero...meaning it was below the 9pg-5pg range that we can currently detect.

We are currently testing know DNA diluted down (serial dilutions) along with other samples of unknown low concentration. We are trying any assay we can think of including Qubit and qPCR. If anyone has any tips or insight it would be greatly appreciated!

[keithsweatshirt]

We haven't used this yet, but are looking into working with it for exactly your reason too.

https://biotium.com/product/accublue-nextgen-dsdna-quantitation-kit/

[Biotium]

Our most sensitive DNA quantitation kit, the AccuBlue NextGen dsDNA Quantitation Kit, has a linear range 0.5-300 pg/uL dsDNA (the kit can detect as low as 0.1 pg/uL on some instruments). This is a 96-well plate assay that uses 10 uL of your DNA sample, it also has been tested in 384 well format with 2.5 uL DNA sample. It requires a fluorescence microplate reader that can read green fluorescence.

Other dsDNA quantitation kits with a variety of linear ranges are also available. Visit our website for more information:

https://biotium.com/technology/accublue-and-accuclear-dsdna-quantitation-kits/