Tweet
Briefly, cDNA from single neurons prepared using SMART-Seq4 kit spent 90 minutes at 95°C due to human error. Is the cDNA to be discarded or can it still be used for library preparation with Nextera XT and sent to sequencing?
Thanks
Thanks
-
-
Prolonged high heat exposure definitely introduces a lot of damages to DNA. I would expect it to be pretty fragmented by now. I also assume the Nextera transpose would require dsDNA - thus, you would need to add at least another PCR cycle. Perhaps you could use this S1 kit instead?: https://swiftbiosci.com/products/acc...-library-kits/
If the sample can be reproduced, best dump it.Last edited by luc; 07-13-2018, 05:55 PM.
-
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 -
With the launch of new single-cell sequencing platforms in 2026, the field stands at an exciting inflection point. This article surveys the most impactful advances in the field and discusses how they’re reshaping research in cancer, immunology, and beyond.
Introduction
Single-cell sequencing technologies have undergone remarkable advances over the past decade, transitioning from low-throughput experimental approaches to highly scalable platforms capable of...05-22-2026, 06:42 AM
Comment