Tweet
We are sending ChIP samples for sequencing on a HiSeq 2500. We sent some to a facility a while ago and they used an Illumina kit for library prep. Now we are planning to switch to a new facility for various reasons including cheaper prices and they use a KAPA library prep kit. We want to repeat the original experiment to have n=2. Will there be a problem, or any kind of bias, or anything else we should worry about with one set being prepared with the Kapa kit and the other with the Illumina kit? Also, are these Kapa kits of comparable quality to the Illumina kits?
-
-
If the libraries are constructed using the same adapter stocks, and amplification is done using the same polymerase, the results should be comparable;
Although the % of input DNA converted to adapter-ligated molecules may differ slightly between the kits, this will be rectified in the amp. However, if different enzymes are used for the enrichment, the enzyme which shows the most bias will produce higher duplication rates i.e. you will have more copies (duplicates) of the "easy" fragments, and fewer duplicates of the "difficult" molecules.
-
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