Hi, You can use the instruction in your preparation kit. If you do not use any kits, see the instruction of your manual then homogenate the sample in the extraction buffer. If it is confusing, please send me your manual.
Best, A.
Hello you would have to isolate and homogenize the antennae tissue. You can probably use one of Qiagen's kits ti do the extraction. Magnetic bullet blenders or Omni Homogenizers could be used with Phenol/Chloroform to extract the RNA.
If you want to investigate expression patterns you would then have to run a Taqman Experiment on the Extracted RNA using known genes that express in the antennae.
A simple way would be to freeze your sample in liquid nitrogen and grind up the frozen tissue. A 1.5 ml tube and a pestle works well for this. Add Trizol reagent to the sample. store at -80C.
Your simple should be to freeze your sample in liquid nitrogen and grind up the frozen tissue you can use the following materials
Chloroform;
Isopropanol;
75% and 70% ethanol (prepared with DEPC-treated water);
TRIzol (1ml);
50-100mg samples;
50 ml Screw Cap tube or centrifuge tube;
Chloroform (1/5 volume of trizol);
RNase free water (filtered or DEPC).
1-add chloroform (1/5 volume of trizol; e.g. 0.2ml to 1ml);
2-shake for 15 sec (Eccles protocol: do not vortex);
3-incubate 2-5 min at RT;
4-spin max. 12000g, 5-15 min, 2-8°C;
5-transfer aqueous upper phase into new tube
* RNA precipitation and wash (20-40 min depending on number of samples
* D.RNA wash (15-30 min depending on number of samples)
1-wash pellet with 70% EtOH
*redissolving of RNA
1-dissolve pellet in 50-100 µl filtered or DEPC H2O (note: DEPC inhibits RT reaction);
2-pipetting up and down;
3- and heat to 55-60°C for 10 min
Good luck
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