Tweet
Hello all. Can anyone offer insight into the pros and cons of placing index barcodes at the 5' versus the 3' end of the read?
-
-
5' of the read allows you to run multiplexed samples without doing the index read. I liked to do this on the GAII to avoid using the Paired-End module for SR multiplexing. Con is that the bases need to be balanced or you can throw off the cluster calling algorithms. Use cycle 5+ to avoid this type of problem.
3' end of read must mean the standard Illumina indexing method using a second priming and read to hit the index. That method works fine, extra reagents are in the SBS kits to allow for 7 cycles for the index read. Illumina only has 12-indexes for DNA and RNA out so far, so that would be a con. On the HiSeq this indexing read is easy to set up, I avoided it on the GAII. -
Thanks for the info!Comment
-
Bioo has 48 barcodes using the 3' method. The kit works well.Comment
-
The field of epigenetics has traditionally concentrated more on DNA and how changes like methylation and phosphorylation of histones impact gene expression and regulation. However, our increased understanding of RNA modifications and their importance in cellular processes has led to a rise in epitranscriptomics research. “Epitranscriptomics brings together the concepts of epigenetics and gene expression,” explained Adrien Leger, PhD, Principal Research Scientist...Yesterday, 07:01 AM -
Proteins are often described as the workhorses of the cell, and identifying their sequences is key to understanding their role in biological processes and disease. Currently, the most common technique used to determine protein sequences is mass spectrometry. While still a valuable tool, mass spectrometry faces several limitations and requires a highly experienced scientist familiar with the equipment to operate it. Additionally, other proteomic methods, like affinity assays, are constrained...04-04-2024, 04:25 PM
Comment