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Does anyone know the composition of the Tagment Wash Buffer in the Nextera DNA Flex Library Prep (20018704) protocol? Does it contain alcohol? PEG?
TIA
TIA
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I would check out the SMART-SEQ2 protocol. They did homebrew something similar. I vaguely remember SDS to denature the enzyme. -
Hi BexMultiplex,
According to the Nextera Flex patent, TWB is:
-100 mM Tris HCl pH 8.5
-100 mM NaCL
-0.1% Tween 20
We have a Tecan, and are going to be reducing RXN volumes down to 1/4, and at that point TWB becomes a limiting reagent. I will be trying my own eventually, but I have not used it yet personally myself, so i can't promise it will work
If you are ever trying to find what a reagent is, looking at the patent (if you can find it) is probably the best way to get this kind of information. It also really helpful for understanding how a kit work when the protocol is vague.
Hope this helps!Comment
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Thanks @UCan'tBcereus, that's really helpful!Comment
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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...06-18-2026, 07:11 AM -
Data variability is still an issue in sequencing technologies despite the advances in reproducibility and accuracy of these platforms. But the problem does not originate in the sequencing itself, but in the previous steps, before the sample reaches the sequencer.
The first step is collection, followed by preservation and sample preparation for analysis. Most scientists overlook those steps, but not being careful might just be skewing the experiment’s results.
...06-02-2026, 10:05 AM
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