Absolutely RNA FFPE Kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	- If RNA degradation is high, ensure that Proteinase K digestion is carried out at 55°C. If RNA yield is poor ensure that the 90% sulfolane and the pre-filter spin cup filtrate were combined at a 1:1 ratio prior to loading the RNAbinding spin cup.	- If Final RNA concentration is too low for use in subsequent applications use a smaller volume of Elution Buffer, ensuring that the surface of the fiber matrix is completely covered.

Protocol tips
- If RNA degradation is high, ensure that Proteinase K digestion is carried out at 55°C. If RNA yield is poor ensure that the 90% sulfolane and the pre-filter spin cup filtrate were combined at a 1:1 ratio prior to loading the RNAbinding spin cup.
Downstream tips
- If Final RNA concentration is too low for use in subsequent applications use a smaller volume of Elution Buffer, ensuring that the surface of the fiber matrix is completely covered.

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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Manufacturer protocol

Download the product protocol from Agilent Technologies for Absolutely RNA FFPE Kit below.

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