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Can somebody help me in getting the new RL protocol compatible with the Nimblegen capture downstream? I tried to design the old Titanium adaptors myself with a T overhang but they didn't work obviously. Got no PCR product. Has somebody ever tried that approach successfully? Perhaps I made a mistake during oligo/adaptor design??? Thanks so much. Tom
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I think the primers you would normally use for you LM-PCR work on the rapid library as well. The biggest problem I see is that your blockers (Hybridization Enhancing Oligo's) won't work as well as they do for general libraries so if this would effect your ontarget you'll have to design them. Perhaps even per MID.
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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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I’m not a sequencing expert. I’m a purification scientist who uses NGS to evaluate workflows my group develops. With this perspective, we think about the sample first and the NGS workflow second. The sequencer is an exceptionally honest reporter, but it can only report on what you give it, so whether you get clean, interpretable data from an NGS workflow is largely determined before you begin.
Here are nine questions we think about, in roughly the order they matter, before...
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