How does the smaller number of UMI (144) provide a complete diversity of tagged molecules in ThruPLEX Tag-seq kits?
How does the smaller number of UMI (144) provide a complete diversity of tagged molecules in ThruPLEX Tag-seq kits?
First of all, there are no identical fragments with the exact same shearing pattern or same extremities at both 5’ and 3’ ends. So, we can say there is extremely low chance of having identical fragment with the same ends.
Secondly, there is extremely low chance of ligation of the same UMI combination to both ends of a fragment even there were identical fragments.
Thirdly, the diversity comes from the orientation of ligation as both ends of DNA equally accessible to both - P5 and P7. If there were identical fragments, the P5 and P7 flip ends adding another level of diversity. " "AssayPrinciple, ProdDescription"
What Bioinformatics tools do you recommend for the PicoPLEX Gold DNA-seq kits? "We do not recommend any particular software tools for PicoPLEX Gold DNA-seq kits.
Though there are several open source and commercially available software packages.
A choice of Software depends on specific customer needs and each Software may offer certain features.
We have used two in house,
• CNV-seq - (http://tiger.dbs.nus.edu.sg/cnv-seq/) and
• BioDiscovery Nexus which is compatible with data collected from the Illumina MiSeq and provide CNV analysis (http://www.biodiscovery.com/nexus-copy-number/)