Trizol RNA extraction of bodies, Flychip Protocol

Published on Author Anne Petzold


"Small scale extraction of total RNA from Drosophila melanogaster" @

(extraction based on combination of Trizol and Chloroform)


app. 30 male fly bodies.


After incubation @ -20 DEG (step 12), sample was split in two. Sample 1 was temporarily stored @ -2 DEG in 500 ul ethanol, while sample 2 was treated with DNase I.

DNase I treatment (following manufacturer's instructions):

- resuspend 10 ug RNA in 1x DNase I reaction buffer to final volume of 100ul

- add 0.5 ul of DNase I, mix, incubate @ 37 DEG for 10"

- add 1ul 0.5 M EDTA (microfiltered and autoclaved EDTA)

- heat inactivate @ 75 DEG for 10 "

- resume original protocol, step 12 (spin down RNA)


Sample 1 (no DNase I treatment): [260/280] 1.75, [260/230] 0.73, 489.7 ug/ul (Nanodrop, RNA-40 mode)

Indicative of contamination with phenol or protein - can I still use it? How to best store it? Now stored @ -20 DEG. Probable sources of contamination: #1 pestle, other: vortex longer when chloroform extracting to get rid of proteins, homogenize more, here some tips:

Sample 2 - did not produce a pellet after step 12, discarded