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Team:SDU-Denmark/labwork/Notebook
From 2012.igem.org
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| - | The procedure used to isolate mRNA from | + | The procedure used to isolate mRNA from Helianthus tuberosus, was a modified version of the Qiagen RNeasy protocol for plant mRNA isolation. See <a href="https://2012.igem.org/Team:SDU-Denmark/labwork/Protocols"><b>mRNA isolation</b></a> protocol for details<br/> |
Plant material form <i>Helianthus tuberosus</i> 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. <br/> | Plant material form <i>Helianthus tuberosus</i> 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. <br/> | ||
| - | 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. <br/> | + | 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 it, and to further disrupt the cell wall. <br/> |
| - | After | + | After centrifugation, Ethanol was added to the solution and the sample was transfered to a spin-column from the Qiagen RNeasy mini kit. The kit i usually used for RNA isolation from animal cells was used (see protocol for details). |
| - | The now isolated mRNA solutions were | + | The now isolated mRNA solutions were measured on a <b>Nanodrop</b> to ascertain their concentrations: |
<p> | <p> | ||
Control: 627 ng/uL <br/> | Control: 627 ng/uL <br/> | ||
Revision as of 16:17, 25 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:
Plant material from the Jerusalem artichoke (Helianthus tuberosus) were secured from a nearby plant nursery(Langeskov Planteskole). The
The procedure used to isolate mRNA from Helianthus tuberosus, was 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 it, and to further disrupt the cell wall.
After centrifugation, Ethanol was added to the solution and the sample was transfered to a spin-column from the Qiagen RNeasy mini kit. The kit i usually used for RNA isolation from animal cells was used (see protocol for details).
The now isolated mRNA solutions were measured on a Nanodrop to ascertain their concentrations:
Control: 627 ng/uL
Sonic: 747 ng/uL
