Leukocyte RNA Purification Kit

Leukocyte RNA Purification Kit

Catalog# : NB-21200
Price/Unit : $295.00
Unit : 50 preps

For the rapid extraction and purification of total RNA from leukocytes

* Fractionate leukocytes from whole blood in minutes
* Isolate total RNA, including microRNA, without phenol
* Rapid and convenient spin-column format

Bio-Synthesis's Leukocyte RNA Purification Kit provides a rapid method for the isolation and purification of total leukocyte RNA from mammalian blood samples in 40 minutes. Selective isolation of leukocyte RNA results in improved expression profiling and other downstream applications by removing the masking effects of some RNAs which are very abundant in whole blood, such as globin mRNAs. The kit is able to isolate total leukocyte RNA, including both large mRNA and all small RNA species containing microRNA (miRNA) and small silencing RNA (siRNA). The purified RNA is of the highest quality and can be used in a number of downstream applications.

Purification is based on spin column chromatography using Bio-Synthesis's proprietary resin as the separation matrix. Briefly, the red blood cells are first removed from the sample through differential red blood cell lysis, and the leukocytes are recovered through centrifugation. The leukocytes are lysed, and the leukocyte RNA is bound to Bio-Synthesis's column (BIND). Under these conditions only the RNA will bind to Bio-Synthesis's resin, while the DNA, proteins and other contaminants will be removed in the flowthrough. The bound RNA is then washed to remove any remaining impurities (WASH). Lastly, the purified RNA will be eluted into 50 µL of the provided Elution Buffer or water (ELUTE). Please see the procedure flowchart to the right.

FEATURES AND BENEFITS

* Fast and easy processing - Rapid spin-column format allows for the processing of 10 samples in 40 minutes.
* No phenol:chloroform extractions - Bio-Synthesis's Leukocyte RNA Purification Kit isolates RNA without the use of harmful chemicals such as phenol or chloroform.
* Recovered RNA is suitable for downstream applications - Purified RNA 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 analysis requiring the use of intact RNA.
* Isolate total leukocyte RNA - All leukocyte RNA species are isolated, from large mRNA down to microRNA.
* Fractionate leukocytes from whole blood in minutes - Rapid removal of red blood cells from whole blood samples using differential red blood cell lysis.

APPLICATIONS

The purified RNA is of the highest quality and can be used in a number of downstream applications including:

* Bioanalyzer
* Quantitative, real-time RT-PCR for large RNA and small RNA including miRNA
* RT-PCR for large RNA and small RNA including miRNA
* Northern blotting
* RNase protection
* Primer extension
* Expression array assays
* Next Generation Sequencing
* microRNA Cloning

Figure 1. Consistent Isolation of High Quality Leukocyte RNA. Bio-Synthesis's Leukocyte RNA Purification Kit isolates leukocyte RNA of high quality with great consistency. Total leukocyte RNA was isolated from 100 µL of hamster blood using Bio-Synthesis's Leukocyte RNA Purification Kit. A total of 6 replicates were performed, and 7 µL of the 50 µL purified RNA was then resolved on a 1.2% formaldehyde-agarose gel. As it can be seen, Bio-Synthesis's kit not only isolated high and consistent yields of total RNA, but the RNA was also of high quality as evidenced by intactness of the major 28S and 18S rRNA.


Figure 2. Higher Yield and Quality of Leukocyte RNA Isolated by Bio-Synthesis's Leukocyte RNA Purification Kit. Bio-Synthesis's Leukocyte RNA Purification Kit isolates Leukocyte RNA that exceeds the yield and quality of competitors. Total RNA was isolated from 200 µL of hamster blood using Bio-Synthesis's Leukocyte RNA Purification Kit and a leading competitor’s kit. One microliter of the 50 µL purified RNA was resolved on an Agilent RNA Nano 6000 chip. The gel diagram (Upper panel) and the electropherogram (Lower panel) showed better quality of RNA isolated by Bio-Synthesis's kit. In particular, RNA isolated by Bio-Synthesis's Leukocyte RNA Purification Kit did not have any evidence of the RNA degradation that was present in RNA isolated by the competitor’s kit (Red Circle). In addition, Bio-Synthesis's kit isolated higher amounts of RNA (Blue Line) with the additional recovery of small RNAs including miRNA (Black Circle) which were not present in RNA isolated by the competitor’s kit. The above observation was further confirmed in RT-qPCR reactions as illustrated Figure 3.

Figure 3. High Yield of a Diversity of RNA Species. Bio-Synthesis's Leukocyte RNA Purification Kit effectively recovers all sizes of RNA from large mRNA to small RNA including microRNAs. Total RNA was isolated from 200 µL of hamster blood sample using Bio-Synthesis's Leukocyte RNA Purification Kit and a leading competitor’s kit as illustrated in Figure 2. Two hundred nanograms of the purified RNA (both 50 µL elution volumes) was then used as the template in a RT-qPCR for detecting the beta-Actin gene (Lower Panel) and for detecting miR-21(Upper Panel). In both graphs the blue lines correspond to Bio-Synthesis's isolated-RNA and the red lines correspond to competitor-isolated RNA. As it can be seen, Bio-Synthesis's kit isolated higher yields of microRNA, as indicated by the lower Ct values of the blue lines (Upper Panel). Also, Bio-Synthesis's kit successfully isolated a similar amount of the large RNA compared to the competitor’s kit (Lower Panel) indicating the full diversity of RNA species isolated.

Leukocyte RNA Purification Kit Contents

1.RBC Lysis Solution
2.Binding Solution
3.Wash Solution
4.Elution Buffer
5.Micro Spin Columns
6.Collection Tubes
7.Elution Tubes
8.Product Insert

Long-Term Storage : All solutions should be kept tightly sealed and stored at room temperature. The RBC Lysis Buffer should be stored at 4°C upon arrival. These reagents should remain stable for at least 1 year in their unopened containers.
Product Usage
For laboratory research use only, not for diagnostic use! Not for use in humans.