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Total RNA Purification Kit for Leukocytes Add to Cart

Cat#: 21200-NB
Quantity: 50 preps
Price: 370 €
Supplier: Norgen
Shipping: RT
User Manual  

• Fast and easy processing using rapid spin-column format
• No phenol or chloroform extractions
• Differential red blood cell lysis allows for the removal of a majority of globin mRNAs
• Isolate total leukocyte RNA, including all small RNA species
• High quality leukocyte RNA can be used in a number of downstream applications


Norgen´s Leukocyte RNA Purification Kit provides a rapid method for the isolation and purification of total leukocyte (white blood cell) RNA from mammalian blood samples. RNA isolated from blood can be used in various expression studies including those focusing on diseases. However, a major problem with blood RNA isolation is that a large portion of the RNA present is globin mRNA, which is found primarily in red blood cells. In fact, up to 70% of the mass of mRNA in whole blood total RNA is globin transcripts. Therefore, it is desirable to be able to remove the red blood cells from the sample and isolate only the RNA associated with the leukocytes, which will result in improved expression profiling and other applications by removing the masking effects of this abundant globin mRNA. Norgen´s Leukocyte RNA Purification Kit can be used isolate and purify total leukocyte RNA, including all small RNAs, from mammalian blood samples.


Norgen´s Purification Technology

Purification is based on spin column chromatography using Norgen´s proprietary resin as the separation matrix. The RNA is preferentially purified from the other cellular components such proteins, without the use of phenol or chloroform. For leukocyte RNA purification, whole blood samples are first collected with anticoagulants. The red blood cells are removed through differential red blood cell lysis, and the leukocytes are recovered by centrifugation. The recovered leukcoytes are then lysed, and the lysate is loaded onto a supplied spin column. Norgen´s resin binds RNA in a manner that depends on ionic concentrations. Thus only the RNA will bind to the column, while the contaminating proteins and genomic DNA will be removed in the flowthrough or retained on the top of the resin. The bound RNA is then washed twice with the provided wash buffer in order to remove any remaining impurities, and the purified leukocyte RNA is eluted with the elution buffer. Norgen´s kit allows for the isolation of total leukocyte RNA, including all small RNA species. The purified RNA is of the highest quality and can be used in a number of downstream applications including real time PCR, reverse transcription PCR, northern blotting, RNase protection and primer extension, and expression array assays.


Kit Specifications

- Column binding capacity: 50 µg
- Maximum column loading volume: 700 µL
- Size of RNA purified: All sizes, including small RNA (<200 nt)
- Maximum blood input: 500 µL
- Minimum blood input: 10 µL
- Time to complete 10 purifications: 45 minutes
- Average yield from 500 µL human blood: 1.5 µg


Image

Leukocyte total RNA. Leukocyte total RNA was isolated from 250 µL of rabbit blood from 4 different samples (lanes 1 to 4). Twenty µL of each elution was then run on a 1.5% formaldehyde-agarose gel. Lane M is Norgen´s RNA Ladder.


Image


Consistency and Yield vs. Competitor.
Leukocyte total RNA was isolated from 50 µL of human blood.
20 uL out of 50 µL of each elution was then run on a 1.5% formaldehyde-agarose gel. Lane M is Norgen´s RNA Ladder. Note the quality, consistency and yield of Norgen´s kit (Lane 1 to 4) as compared to the competitor (Lane 5 and 6).

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