I am using targeted sequencing designed by Qiagen. I would like to perform a gel run for my samples before measuring them by Qubid and AMPure step. The reason I want to do that is because if I depended ONLY on Qubid I will have a measurement for everything in my sample, while when I run a gel I can see my targeted amplicons based on their size in the gel. Does any one agree with me? if disagree can you tell me why and correct me if I am wrong?
-
-
With a normal ampure cleanup you can get rid of primers. Depending on exactly what you are doing you can do some size selection as well. Typically people will use a bioanalyzer or a tapestation as a final step to QC their library and to quantify it. If you do a gel extraction first you can lose a lot of material and possibly sacrifice the complexity of your library. -
Thank you for your replay. I want to use the gel just to check the (efficiency) of my PCR that is all. I will not cut it or size select. This is becasue when I measure with quibid I get reading that is much higher than after AMPure which is a very tuff experiance for my sample
as they end up with much lower reading after the AMPure. So I do not know if this low reading becasue my PCR was not working well from the begining or a result of the AMPure step.
Comment
-
Ah, if you're just troubleshooting then go for it. Then you can determine if the problem is the PCR or the Ampure cleanup.Comment
-
NGS has revolutionized clinical microbiology by offering new ways to identify and surveil pathogens without prior knowledge of the target. One popular approach is 16S rRNA and ITS amplicon sequencing, which targets specific microbial markers. But studies are increasingly incorporating shotgun metagenomics, a technique that sequences all DNA from a sample without targeting specific microbial markers. This approach allows more detailed taxonomic classification, detection...Today, 06:46 AM -
Tumors contain complex mixtures of cell populations that evolve in response to treatment and host-specific factors. Tissue biopsies provide only static snapshots of a tumor’s state. But what if we could follow that evolution over time using a minimally invasive technique?
Liquid biopsies can detect tumor-derived material circulating in the bloodstream, providing a way to study treatment response, detect emerging resistance, and discover new biomarkers. Circulating...10-14-2026, 09:55 AM
Comment