Absolutely RNA FFPE Kit

RNA isolation / purification Tissue - Rat Kidney

Experiment
RNA isolation / purification Tissue - Rat Kidney
Product
Absolutely RNA FFPE Kit from Agilent Technologies
Manufacturer
Agilent Technologies

Protocol tips

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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Discussion

Discussion

4 years ago

Author: Paul G. Macon United States

RNA isolation from tissue

How do I extract RNA from animal tissue without using liquid nitrogen? I tried the RNA extraction by using the TRIzol reagent and I homogenize the tissue using polytron homogenizer at room temperature for 30secs is this correct?

Discussion

5 years ago

Author: Aaron Stege Netherlands

Problem in phase separation after using serum/plasma kit

I used a serum/plasma kit for my serum samples. After the phase separation the samples should have 3 phases: a colourless aqueous phase, a white interphase and a red organic phase. However, in some of my samples there was no aqueous phase unless I wait for an extended period of time. How can I circumvent this problem?

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Papers

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Manufacturer protocol

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

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