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Originally posted by TonyBrooks
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Thanks. Looks like I should use the HS kit since my DNA is only about 5 ng/ul -
it's pointed out in another thread somewhere but if you are switching between the 2 kits you need to do some comparison tests of your samples using each kit. BR tends to give a higher reading by a high percentage vs the HS in my experience.Originally posted by lailaizhang View Post
I once ran out of the BR dsDNA assay which I had been using to measure the concentration of my RNAseq libraries for cluster concentration calculations and instead used a HS assay in a pinch, did the calculations the same and clustered at "the same" concentration and all my libraries over clustered. I then did a side by side of the libraries with the BR and HS and there was on average a 29% difference between the values given, with the BR being higher. All the libraries were falling in the 40ng/ul range on the HS so they were well with in the BR kit range
I had good consistent clustering results with the BR assay when using it, and only had a problem with it when i switched to HS without making adjustments for the different values given. (<-- unknown to me at the time, or an oversight) I will stick with HS from now on for anything under 100ng/ul, I do feel like it is more accurate with in that range vs the BRLast edited by subxero; 10-01-2015, 04:23 AM.Comment
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Love this - good data definitely starts from good input, and poor input can only give relatively poor data. I particularly like the mention of Nanodrop/absorbance based methods for quantification. It's such a toss up if you'll get an accurate reading or what amounts to a randomly generated number, and a lot of library/sequencing related issues can be traced back to poor quant.07-01-2026, 11:43 AM
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