SEQanswers - View Single Post - Terrible overclustering -- 16S EMP protocol

neko_33 · 12-02-2016, 09:54 PM

Thank you for your kind replies.

We usually quantify libraries with Qubit fluorometer, but at times with qPCR (KAPA Library Quantification Kits). Illumina Co. warns that Biolanalyzer quanitification is not reliable, according to my memory.

Empirically, not so much difference between Qubit and qPCR. For example, when I last did MiSeq run, the concentration calculated based on Qubit result was 1.9±0.1nM, while 2.2nM on qPCR.

As I wrote in the first message, I think our quantifications do not have thus fatal errors, but I will continue using both Qubit and qPCR, and compare these two results.

12-02-2016, 09:54 PM	#4
neko_33 Junior Member Location: Tokyo Join Date: Dec 2016 Posts: 3	quantification failure? -- maybe not Thank you for your kind replies. We usually quantify libraries with Qubit fluorometer, but at times with qPCR (KAPA Library Quantification Kits). Illumina Co. warns that Biolanalyzer quanitification is not reliable, according to my memory. Empirically, not so much difference between Qubit and qPCR. For example, when I last did MiSeq run, the concentration calculated based on Qubit result was 1.9±0.1nM, while 2.2nM on qPCR. As I wrote in the first message, I think our quantifications do not have thus fatal errors, but I will continue using both Qubit and qPCR, and compare these two results.