High-throughput polyribosome fractionation.
Polyribosome sedimentation velocity centrifugation can be used to identify differential regulation of the translation of mRNAs. However, ultracentrifugation presents practical limitations on the number of sedimentation velocity gradients that can be run simultaneously. A method for sedimentation velocity analysis of polyribosomes is presented that is based on low-speed centrifugation of sucrose gradients prepared in deep 96-well plates, the advantage of which is that hundreds of polyribosome fractionations can be performed simultaneously in a tabletop centrifuge.
Wang
Yipeng
Y
Sidney Kimmel Cancer Center, 10835 Altman Row, San Diego, CA 92121, USA.
Ringquist
Steven
S
Cho
Ann H
AH
Rondeau
Gaelle
G
Welsh
John
J
D002458:Cell Fractionation / Q000379:methods*; D045744:Cell Line, Tumor; D002499:Centrifugation, Density Gradient; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D011132:Polyribosomes / Q000187:drug effects / Q000378:metabolism*; D014176:Protein Biosynthesis / Q000187:drug effects; D011691:Puromycin / Q000494:pharmacology; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014461:Ultracentrifugation
D012333:RNA, Messenger; D011691:Puromycin