Protocol tips
Upstream tips |
Protocol tips |
Downstream tips |
- To allow complete penetration by formalin, use tissue samples less than 5 mm thick
|
- Perform all centrifugation steps using a microcentrifuge placed at 15–25°C
|
- Do not exceed fixation time of 24 hours as it results in poor performance in downstream assay
|
Upstream tips |
- To allow complete penetration by formalin, use tissue samples less than 5 mm thick
|
Protocol tips |
- Perform all centrifugation steps using a microcentrifuge placed at 15–25°C
|
Downstream tips |
- Do not exceed fixation time of 24 hours as it results in poor performance in downstream assay
|
Publication protocol
In a first step, seven RNA extraction kits (Table 1) were tested on rat renal tissue. 12 samples were analyzed for each kit: Four samples with one rat whole kidney section, four samples with two whole kidney sections and four samples with 80–101 LCM glomerular cross-sections. All sections had a thickness of 5 lm. Later, the four kits, which yielded the highest amount of RNA extracted from rat whole kidney sections were used to extract RNA from human renal tissue: Six samples were analyzed for each kit: Four samples with two human kidney biopsy sections and two samples with 80–160 LCM glomerular cross-sections. Human samples had a thickness of 10 lm. Each of the seven different kits was performed according to the manufacturer’s instructions under RNAse-free conditions. The volume of eluate was set to 25 lL in the case of the rat tissues and 15 lL in the case of the human tissue
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