Leukocyte RNA Purification Kits For the rapid extraction and purification of total RNA from leukocytes For research use only and NOT intended for in vitro diagnostics. Powered by Bioz Leukocyte RNA Purification Kits For the rapid extraction and purification of total RNA from leukocytes Price USD$339.00 SKU 21200 Format Spin Column Plus High Throughput Size 50 Preps Quantity Overview Details Documentation Citations Overview Fractionate leukocytes from whole blood in minutes with provided RBC Lysis Buffer Isolate total RNA, including microRNA, without phenol Rapid and convenient spin-column format and 96-well plate for high throughput applications Purified RNA is ready for any downstream application including RT-PCR, qRT-PCR, NGS, arrays and more. Find out more information on Norgen's NGS services Purification is based on spin column chromatography that uses Norgen’s proprietary resin separation matrix These kits provide a rapid method for the isolation and purification of total leukocyte (white blood cell) RNA from mammalian blood samples. These kits are supplied with an RBC (red blood cell) Lysis Buffer for selective removal of red blood cells and fractionation of leukocytes by centrifugation. 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. These kits are 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. Leukocyte RNA Purification Kit (Spin Column) This kit provides a rapid method for the isolation and purification of total leukocyte RNA from mammalian blood samples in 40 minutes. Allowable blood input ranges from 10 μL to 2 mL or 3 x 106 Leukocytes Leukocyte RNA Purification Plus Kit (Plus) Norgen’s Leukocyte RNA Purification Plus Kit provides a rapid method for the isolation and purification of total leukocyte (white blood cell) RNA from up to 3 mL of mammalian blood samples. Complete 10 purifications in as little as 40 minutes. Leukocyte RNA Purification 96-Well Kit (High Throughput) Purification is based on 96-well column chromatography using Norgen’s proprietary resin as the separation matrix. Purification can be performed using either a vacuum manifold or centrifugation. 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. Allowable blood input ranges from 10 μL to 1 mL or 1.5 x 106 Leukocytes. Details Supporting Data Previous Figure 1. Consistent Isolation of High Quality Leukocyte RNA. Norgen'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 Norgen'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 can be seen, Norgen'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 Norgen's Leukocyte RNA Purification Kit. Norgen'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 Norgen'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 Norgen's kit. In particular, RNA isolated by Norgen'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, Norgen'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 in Figure 3. Figure 3. High Yield of a Diversity of RNA Species. Norgen'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 Norgen'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 Norgen isolated-RNA and the red lines correspond to competitor-isolated RNA. As it can be seen, Norgen's kit isolated higher yields of microRNA, as indicated by the lower Ct values of the blue lines (Upper Panel). Also, Norgen'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. Figure 4. Norgen's Leukocyte RNA Purification Plus Kit (Cat No. 21250) was used to Isolate RNA from 2.5 mL of Whole Blood Collected from 24 donors, Across 3 Experiments. Donors were tested in duplicate. After the standard protocol failed to produce RNA with sufficient concentration, modifications were made to the procedure and 16 additional donors were tested. 2 batches of 8 additional donors were tested in duplicate with the modified protocol for 32 separate RNA isolations; 32/32 yielded RNA concentration to >100 ng/µL. A single pass of 30 µL of elution buffer was used. (Tessa Wiehle, Stephanie Borel, Andrew Kincaid, John Hedges, Jon Kemppainen, Justin T Brown, Andrew G Hadd, and Gary J Latham. Modifications to RNA Isolation Protocols Meet Requirements for Modern CML Monitoring of BCR-ABL1 Transcript Levels. Asuragen Inc. 2018) Figure 5. Consistent Yield of High Quality Leukocyte RNA with Complete Size Range without the Use of Phenol. Norgen's Leukocyte RNA Purification 96-Well Kit allows for the consistent isolation of high quality RNA, with complete size range from the very large RNA down to small RNA without the use of phenol. Total RNA was isolated from 200 µL aliquots of hamster blood using Norgen's Leukocyte RNA Purification 96-Well Kit in 12 replicates. Five microliters of the 75 µL isolated RNA was resolved on a 1.2% formaldehyde agarose gel (Panel A) and 1 µL of the 75 µL isolated RNA was resolved on an Agilent RNA Nano 6000 Chip in a 2100 Bioanalyzer (Panel B). All 12 replicates showed both high yield and high quality. In addition, all replicates showed effective recovery of all sizes of RNA including the small RNA (arrow). Figure 6. Isolation of Both Large and Small RNA. Norgen's Leukocyte RNA Purification 96-Well Kit allows for consistent isolation of both large and small RNA. Leukocyte RNA was isolated from 200 µL of hamster blood using Norgen's Leukocyte RNA Purification 96-Well Kit. Aliquots of each total RNA sample were then used in 2 different RT-qPCR reactions. Ten microliters of the 75 µL isolated RNA was subjected to a 20 µL reverse transcription using miR-21 stem-loop reverse primer or oligo dT. Three microliters of the reverse transcription was used in a 20 µL real-time PCR reaction with primers to detect the miR-21 (Panel A) and the beta-actin transcripts (Panel B), respectively. Both the mRNA and microRNA were amplified in a consistent manner from all the samples, with low variability of the Ct values. Thus both types of RNA are being consistently isolated using this kit. Next Click for expanded view Kit Specifications Maximum Column Binding Capacity Up to 50 μg RNA Maximum Loading Volume 650 μL Size of RNA Purified All sizes, including small RNA <200 nt Minimum Blood Input 10 μL Maximum Blood Input 2 mL or 3 x 106 leukocytes Time to Complete 10 Purifications 40 minutes Input Type Blood from Human, Hamster, Mouse, Rabbit, and Rat Average Yield Human Blood (500 μL) Hamster Blood (100 μL) 1.5 μg 2.5 μg Storage Conditions and Product Stability All solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 2 years after the date of shipment. The RBC Lysis Buffer should be stored at 4°C upon arrival. Component Cat. 21200 (50 preps) Cat. 21250 (50 preps) Cat. 37800 (2 x 96 preps) RBC Lysis Buffer 2 x 100 mL 2 x 1 L 2 x 90 mL Buffer RL 30 mL 30 mL - Binding Solution - - 2 x 40 mL Wash Solution A 38 mL 38 mL - Wash Solution - - 2 x 30 mL Elution Solution A 6 mL 6 mL - Elution Solution - - 2 x 20 mL Enzyme Incubation Buffer - 6 mL 1 DNase I - 1 Vial - Mini Spin Columns 50 - - Lysate Homogenization Column - 50 - Single Cell RNA Column - 50 - RBC Lysis 96-Well Plate - - 2 96-Well Filter Plate - - 2 Adhesive Tape - - 4 Collection Tubes 50 100 - 96-Well Collection Plate - - 2 Elution Tubes (1.7 mL) 50 50 - 96-Well Elution Plate - - 2 Product Insert 1 1 1 Documentation PROTOCOLS (21200) Leukocyte RNA Purification Kit (Spin Column) - Protocol (50 prep) (21250) Leukocyte RNA Purification Plus Kit (Plus) - Protocol (50 prep) (37800) Leukocyte RNA Purification 96-Well Kit (High Throughput) - Protocol (96-well) APPLICATION NOTES Rapid Isolation and Purification of Total Leukocyte White Blood Cell RNA From Mammalian Blood Samples SAFETY DATA SHEETS (21200, 21250) Leukocyte RNA Purification Kit - Safety Data Sheet (37800) Leukocyte RNA Purification 96-Well Kit - Safety Data Sheet RELATED BLOGS 3 Reasons why Norgen’s Industry-Leading Patented RNA Purification Technology is Best-in-Class How to Reduce False Negative SARS-CoV-2 Results with Silicon Carbide Citations Spin Column (21200) Title Effects of Omega-3 Supplementation on Components of the Endocannabinoid System and Metabolic and Inflammatory Responses in Adipose and Liver of Peripartum Dairy Cows Journal Research Square. 2022. Authors Gitit Kra, Jayasimha Rayalu Daddam, Uzi Moallem, Hadar Kamer, Radka Kočvarová, Alina Nemirovski, G. Andres Contreras, Joseph Tam and Maya Zachut Title MicroRNA molecular profiling from matched tumor and bio-fluids in bladder cancer Journal Molecular Cancer. 2015. Authors David A. Armstrong, Benjamin B. Green, John D. Seigne, Alan R. Schned and Carmen J. Marsit Title Novel Noncompetitive IL-1 Receptor-Biased Ligand Prevents Infection- and Inflammation-Induced Preterm Birth Journal Journal of Immunology. 2015. Authors Nadeau-Vallée M, Quiniou C, Palacios J, Hou X, Erfani A, Madaan A, Sanchez M, Leimert K, Boudreault A, Duhamel F, Rivera JC, Zhu T, Noueihed B, Robertson SA, Ni X, Olson DM, Lubell W, Girard S, Chemtob S Title Enhanced antioxidant capacity and anti-ageing biomarkers after diet micronutrient supplementation. Journal Molecules. 2014. Authors Balcerczyk, Gajewska, Macierzyńska-Piotrowska, Pawelczyk, Bartosz, Szemraj. Title Increased TNF α, IL-6 and ErbB2 mRNA expression in peripheral blood leukocytes from breast cancer patients. Journal Med Oncol. 2014. Authors Alokail MS, Al-Daghri NM, Mohammed AK, Vanhoutte P, Alenad A. Title Increased Il-4 Mrna expression and poly-aromatic hydrocarbon concentrations from children with asthma. Journal BMC Pediatrics. 2014. Authors Al-Daghri NM, Abd-Alrahman S, Draz H, Alkharfy K, Mohammed AK, Clerici MS, Alokail MS. Title Targeted Inhibition of Serotonin Type 7 (5-HT7) Receptor Function Modulates Immune Responses and Reduces the Severity of Intestinal Inflammation. Journal Journal of Immunology. 2013. Authors Kim JJ, Bridle BW, Ghia JE, Wang H, Syed SN, Manocha MM, Rengasamy P, Shajib MS, Wan Y, Hedlund PB, Khan WI. Title Biodosimetry on small blood volume using gene expression assay. Journal Health Physics, 98(2):179-85. 2010. Authors Brengues M, Paap B, Bittner M, Amundson S, Seligmann B, Korn R, Lenigk R, Zenhausern F. Plus (21250) Title MicroRNA molecular profiling from matched tumor and bio-fluids in bladder cancer Journal Molecular Cancer. 2015. Authors David A. Armstrong, Benjamin B. Green, John D. Seigne, Alan R. Schned and Carmen J. Marsit Title Novel Noncompetitive IL-1 Receptor-Biased Ligand Prevents Infection- and Inflammation-Induced Preterm Birth Journal Journal of Immunology. 2015. Authors Nadeau-Vallée M, Quiniou C, Palacios J, Hou X, Erfani A, Madaan A, Sanchez M, Leimert K, Boudreault A, Duhamel F, Rivera JC, Zhu T, Noueihed B, Robertson SA, Ni X, Olson DM, Lubell W, Girard S, Chemtob S Title Enhanced antioxidant capacity and anti-ageing biomarkers after diet micronutrient supplementation. Journal Molecules. 2014. Authors Balcerczyk, Gajewska, Macierzyńska-Piotrowska, Pawelczyk, Bartosz, Szemraj. Title Increased TNF α, IL-6 and ErbB2 mRNA expression in peripheral blood leukocytes from breast cancer patients. Journal Med Oncol. 2014. Authors Alokail MS, Al-Daghri NM, Mohammed AK, Vanhoutte P, Alenad A. Title Increased Il-4 Mrna expression and poly-aromatic hydrocarbon concentrations from children with asthma. Journal BMC Pediatrics. 2014. Authors Al-Daghri NM, Abd-Alrahman S, Draz H, Alkharfy K, Mohammed AK, Clerici MS, Alokail MS. Title Targeted Inhibition of Serotonin Type 7 (5-HT7) Receptor Function Modulates Immune Responses and Reduces the Severity of Intestinal Inflammation. Journal Journal of Immunology. 2013. Authors Kim JJ, Bridle BW, Ghia JE, Wang H, Syed SN, Manocha MM, Rengasamy P, Shajib MS, Wan Y, Hedlund PB, Khan WI. Title Biodosimetry on small blood volume using gene expression assay. Journal Health Physics, 98(2):179-85. 2010. Authors Brengues M, Paap B, Bittner M, Amundson S, Seligmann B, Korn R, Lenigk R, Zenhausern F. Related Services NGS RNA Isolation Related Products RNase-Free DNase I Kit Cytoplasmic & Nuclear RNA Purification Kit microRNA Purification Kit
Leukocyte RNA Purification Kits For the rapid extraction and purification of total RNA from leukocytes For research use only and NOT intended for in vitro diagnostics. Powered by Bioz Leukocyte RNA Purification Kits For the rapid extraction and purification of total RNA from leukocytes Price USD$339.00 SKU 21200 Format Spin Column Plus High Throughput Size 50 Preps Quantity