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Revision as of 16:34, 6 September 2012
Laboratory Notebook
Here you find the log book for the procedures carried out in the laboratory, starting from week 27.05-07-2012:
Securing plant material
Plant material from Jerusalem artichoke (Helianthus tuberosus) were secured from a nearby plant nursery (Langeskov Planteskole).
06-07-2012:
Isolation of mRNA from Helianthus tuberosus
(The procedure we used for mRNA isolation, is a modified version of the Qiagen RNeasy protocol for plant mRNA isolation. See (mRNA isolation) protocol for details)
Plant material form Helianthus tuberosus was cut into pieces and stored in liquid nitrogen(flash-freeze at -196 °C)to halt all RNase activity. RLT buffer is mixed at this point, for later use.
After 20 minutes of freezing time the plant material was grinded into fine dust and dissolved in RTL buffer. The solution was treated with ultrasound to homogenize and further disrupt the cell wall.
After centrifuging the solution and adding ethanol to the sample was transfered to a spin-column and Qiagen RNeasy mini kit, for RNA isolation from animal cells was used (see protocol for details).
The now isolated mRNA solutions were tested on Nanodrop:
Control: 627 ng/uL
Sonic: 747 ng/uL
As expected, disruption of the cell wall by ultrasound resulted in a higher yield.
