Next, 10 μl of RT mix was added (and mixed via pipette trituration) to 10 μl of the standardised mRNA solution.
Approximately, 10 μL of cDNA synthesis mix was added to each RNA/primer mixture, gently mixed, briefly centrifuged and incubated at 50 °C for 50 min.
RNAfectin mix was added to the siRNA tube and gently mixed.
250 µl of the siRNA TransIT-siQUEST mix wasiRNA TransIT-siQUESTcontaining 1.25 mixof media washout addediotocs.
Cell pellets were then re-suspended in 25-ml cold PBS, and 2 ml EB/AO (1 1, 4 mg/ml) dye mix was added.
For each gram of calcinated sample, 4 ml of acid mix was added in order to eliminate impurities (calcium, potassium, magnesium, manganese, iron, boron and phosphorous).
Nine and a half micro-litres of master mix was added to each well.
The reaction mix was added to 100 ng of template DNA.
The transfection mix was added drop-wise onto the cells which were incubated for 72 h.
The Matrigel mix was added with 1% antiserum against BDNF or NSS.
One milliliter of this reagent mix was added to the homogenized tissue, followed immediately by 0.2 ml of chloroform.
