How much tissue do I need?
We require 40-50 µg total RNA per direct labelling reaction per slide, i.e. ≈200 µg total RNA per sample. However, we can also perform labelling reactions with less total RNA if an amplification step is included. Where possible, it is advisable to avoid amplification as this is a potential source of systematic bias within the expression ratio measurements.
Please ensure that you have a sufficient amount of material before sending us your samples. Growth conditions, physiological state and mutant phenotype will affect RNA yield, because these will affect both the growth and development of the organism. If you require further assistence or advice, please contact flychip.
We would be happy to perform a pilot study to define how much total RNA can be extracted from your experimental tissue.
Direct labelling without amplification (≈50 µg total RNA):
| Tissue type | Estimated mass (mg) | Estimated quantity |
|---|---|---|
| S2 cells | - | 8 x 106 |
| Embryos | 10 | 1000 embryos |
| Larvae | 14 | - |
| Adult - male | 30 | 36 flies |
| Adult - female | 30 | 36 flies |
| Adult heads | - | 120 heads |
|
Ovary |
- | 50 pairs |
| Testis | - | 84 pairs |
Direct labelling with amplification (≈1 µg total RNA):
| Tissue type | Estimated quantity |
|---|---|
| Embryos | 10 |
|
Ovary pairs |
5 |
| Testis pairs | 5 |
| Wing discs | 10 |
