Tweet
Has anyone had problems with low RNA recovery after chemical fragmentation for mRNA seq? A collaborator says that he loses ~90% of his material when he fragments chemically. He says that covaris shearing of RNA provides a much greater yield. Has anyone else had the same experience?
-
-
Illumina kits uses chemical fragmentation for mRNA seq and they didn't see any problem with the yield of and so i don't think it is chemical fragmentation leads to low yield. Try to adjust the time of the fragmentation (typically 5 minutes). -
What kit was used for chemical fragmentation?
I used a commercial kit for fragmentation. The time was 15 mins. I tried fragmenting for 10 mins and increased it by 1 min up to 15 mins. I got the same result, all the RNA had been fragmented to below 100bp. I won't use this kit again.Comment
-
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 -
Despite advancements in sequencing platforms and related sample preparation technologies, certain sample types continue to present significant challenges that can compromise sequencing results. Pedro Echave, Senior Manager of the Global Business Segment at Revvity, explained that the success of a sequencing experiment ultimately depends on the amount and integrity of the nucleic acid template (RNA or DNA) obtained from a sample. “The better the quality of the nucleic acid isolated...03-22-2024, 06:39 AM
Comment