The preparation of poly(A)+mRNA from the hagfish slime gland.
The isolation of translatable poly(A)+mRNA from the slime glands of the Pacific hagfish, Eptatretus stouti, is not possible by the commonly used procedures because of the viscous slime that is formed when the contents of the glands are hydrated. This paper reports on a procedure developed to overcome this problem. Briefly, the tissue was powdered in liquid nitrogen, mixed with sodium lauroylsarcosine and proteinase K and lyophilized. The lyophilized powder was then mixed with 0.3 mm diameter glass beads, thoroughly ground and wetted with buffer and digested at 37 degrees C. The RNA from the digest was recovered by ultracentrifugation through a CsCl cushion. Further purification of the RNA was accomplished by the usual methods with slight modifications.
D000818:Animals; D004591:Electrophoresis, Polyacrylamide Gel; D005088:Exocrine Glands / Q000032:analysis*; D005399:Fishes / Q000378:metabolism*; D005612:Freeze Drying; D006196:Hagfishes / Q000378:metabolism*; D011061:Poly A / Q000302:isolation & purification*; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000302:isolation & purification*; D014783:Viscosity
D012333:RNA, Messenger; D011061:Poly A