Preserved Blood RNA Purification Kit I (for use with Tempus Blood RNA Tubes)
Rapid isolation and purification of total RNA from blood that has been preserved using Tempus™ Blood RNA Tubes
For research use only and NOT intended for in vitro diagnostics.
Preserved Blood RNA Purification Kit I (for use with Tempus Blood RNA Tubes)
Rapid isolation and purification of total RNA from blood that has been preserved using Tempus™ Blood RNA Tubes
Overview
- Compatible with Tempus™ Blood RNA Tubes
- Isolate true total RNA including siRNA and microRNA
- No phenol extractions
- Purify high-quality RNA in 30 minutes
- Purified RNA is suitable for a variety of downstream applications, including Small RNA Sequencing. Find out more information on Norgen's NGS services
- Purification is based on spin column chromatography that uses Norgen’s proprietary resin separation matrix
Norgen’s Preserved Blood RNA Purification Kit I provides a rapid method for the isolation and purification of total RNA from blood that has been preserved using Tempus™ Blood RNA Tubes*. The kit purifies all sizes of RNA, from large mRNA and ribosomal RNA down to microRNA (miRNA) and small interfering RNA (siRNA). The RNA is preferentially purified from other cellular components such as proteins, without the use of phenol or chloroform.
Purification is based on spin column chromatography using Norgen’s proprietary resin as the separation matrix. Norgen’s proprietary resin provides superior affinity to the full size range of RNA molecules, resulting in large and small RNA (miRNA) purification with better linearity and sensitivity. The purified RNA is of the highest integrity, and can be used in a number of downstream applications including real time RT-PCR, RT-PCR, Northern blotting, RNase protection and primer extension, expression profiling, miRNA cloning and amplification and Next Generation Sequencing.
* Tempus™ is a registered trademark of Life Technologies. Tempus™ Blood RNA Tubes are not provided.
Details
Supporting Data
Kit Specifications
|
|
Maximum Column Binding Capacity
|
50 μg
|
Maximum Column Loading Volume |
650 μL
|
Size of RNA Purified |
All sizes, including
small RNA (< 200 nt) |
Time to Complete 10 Purifications |
20 - 30 minutes
|
Average Yield |
5 - 25 μg per 3 mL
preserved human blood |
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.
Component | Cat. 43400 (50 preps) |
---|---|
Tempus™ Blood RNA Tube Diluent | 2 x 90 mL |
Lysis Solution | 40 mL |
Wash Solution | 22 mL |
Elution Solution | 6 mL |
Mini Spin Columns | 50 |
Collection Tubes | 50 |
Elution Tubes (1.7 mL) | 50 |
Product Insert | 1 |
Documentation
FAQs
Spin Column
- Incomplete lysis of blood.
Ensure that the appropriate amount of Lysis Solution was used.
- An alternative elution solution was used.
It is recommended that the Elution Solution supplied with this kit be used for maximum RNA recovery.
- Ethanol was not added to the lysate.
Ensure that the appropriate amount of ethanol is added to the lysate before binding to the column.
- Ethanol was not added to the Wash Solution.
Ensure that 50 mL of 95% ethanol is added to the supplied Wash Solution prior to use.
- Insufficient centrifugation time and speed when pelleting the stabilized blood RNA before column purification.
Centrifuge the samples for at least 30 minutes or longer at 4,000 x g or higher. The RNA pellet was lost during decanting Pour off the supernatant slowly and carefully.
- Insufficient solubilization of blood.
Ensure that the appropriate amount of Lysis Buffer was used.
- Centrifuge temperature is too low.
Ensure that the centrifuge remains at room temperature throughout the procedure. Temperatures below 15°C may cause precipitates to form that can cause the columns to clog.
- RNase contamination.
RNases may be introduced during the use of the kit. Ensure proper procedures are followed when working with RNA. Please refer to “Working with RNA” at the beginning of the protocol's user guide.
- Procedure not performed quickly enough.
In order to maintain the integrity of the RNA, it is important that the procedure be performed quickly.
- Improper storage of the purified RNA.
For short term storage RNA samples may be stored at -20°C for a few days. It is recommended that samples be stored at -70°C for longer term storage.
- RNA was not washed 3 times with the provided Wash Solution A.
Traces of salt from the binding step may remain in the sample if the well is not washed 3 times with Wash Solution A. Salt may interfere with downstream applications, and thus must be washed from the well.
- Ethanol carryover.
Ensure that the dry spin under Column Wash in the centrifugation protocol or the extended vacuum in the vacuum protocol is performed in order to remove traces of ethanol prior to elution. Ethanol is known to interfere with many downstream applications.
Citations
Title | Correlation between gene expression and MRI STIR signals in patients with chronic low back pain and Modic changes indicates immune involvement |
Journal | Scientific Reports . 2022. |
Authors | Maria Dehli Vigeland, Siri Tennebø Flåm, Magnus Dehli Vigeland, Ansgar Espeland, Per Martin Kristoffersen, Nils Vetti, Monica Wigemyr, Lars Christian Haugli Bråten, Elisabeth Gjefsen, Elina Iordanova Schistad, et al. |
Title | The first horse gut microbiome gene catalog reveals that rare microbiome ensures better cardiovascular fitness in endurance horses |
Journal | bioRxiv. 2022. |
Authors | Núria Mach, Cédric Midoux, Sébastien Leclercq, Samuel Pennarun, Laurence Le Moyec, Olivier Rué, Céline Robert, Guillaume Sallé, Eric Barrey |
Title | The LIFE Child study: a population-based perinatal and pediatric cohort in Germany |
Journal | European Journal of Epidemiology. 2017. |
Authors | Poulain T, Baber R, Vogel M, Pietzner D, Kirsten T, Jurkutat A, Hiemisch A,, Hilbert A,, Kratzsch J, Thiery J, Fuchs M, Hirsch C, Rauscher FG, Loeffler M,, Körner A, Nüchter M, Kiess W |
Title | Comparison of blood RNA isolation methods from samples stabilized in Tempus tubes and stored at a large human biobank |
Journal | BMC Research Notes. 2016. |
Authors | Aarem J, Brunborg G, Aas KK, Harbak K, Taipale MM, Magnus P, Knudsen GP, Duale N. |
Title | Evaluation of Existing Methods for Human Blood mRNA Isolation and Analysis for Large Studies |
Journal | PLoS One. 2016. |
Authors | Meyer A, Paroni F, Günther K, Dharmadhikari G, Ahrens W, Kelm S, Maedler K |
Title | Differences in Whole Blood Gene Expression Associated with Infection Time-Course and Extent of Fetal Mortality in a Reproductive Model of Type 2 Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) Infection |
Journal | PLOS ONE. 2016. |
Authors | Wilkinson JM, Ladinig A, Bao H, Kommadath A, Stothard P, Lunney JK, Harding JC, Plastow GS |
Title | Post-weaning blood transcriptomic differences between Yorkshire pigs divergently selected for residual feed intake |
Journal | BMC Genomics. 2016. |
Authors | Liu H, Nguyen YT, Nettleton D, Dekkers JC, Tuggle CK |
Title | miRNA expression profiles and retinal blood vessel calibers are associated with short-term particulate matter air pollution exposure |
Journal | Environmental Research. 2016. |
Authors | Louwies T, Vuegen C, Panis LI, Cox B, Vrijens K, Nawrot TS, De Boever P |
Title | DICER1 and microRNA regulation in post-traumatic stress disorder with comorbid depression |
Journal | Nature Communications. 2015. |
Authors | Aliza P. Wingo, Lynn M. Almli, Jennifer J. Stevens, Torsten Klengel, Monica Uddin, Yujing Li, Angela C. Bustamante, Adriana Lori, Nastassja Koen, Dan J. Stein, Alicia K. Smith, Allison E. Aiello, Karestan C. Koenen, Derek E. Wildman, Sandro Galea, Bekh Bradley, Elisabeth B. Binder, Peng Jin, Greg Gibson & Kerry J. Ressler |
Title | Genetic variants in the DNA repair gene NEIL3 and the risk of myocardial infarction in a nested case–control study. The HUNT Study |
Journal | DNA Repair. 2015. |
Authors | Tonje Skarpengland, Lars Erik Laugsand, Imre Janszky, Luisa Luna, Bente Halvorsen, Carl G.P. Platou, Wei Wang, Lars J. Vatten, Jan Kristian Damås, Pål Aukrust, Magnar Bjørås, Bjørn O. Åsvold |
Title | Comparison of Whole Blood RNA Preservation Tubes and Novel Generation RNA Extraction Kits for Analysis of mRNA and MiRNA Profiles |
Journal | PLOS ONE. 2014. |
Authors | Madlen Häntzsch, Alexander Tolios, Frank Beutner, Dorothea Nagel, Joachim Thiery, Daniel Teupser, Lesca M. Holdt. |
Title | Anti-Interferon Beta Antibody Titers Strongly Correlate Between Two Bioassays and In Vivo Biomarker Expression, and Indicates That a Titer of 150 TRU/mL Is a Biologically Functional Cut-Point. |
Journal | Journal of Interferon and Cytokine Research. 2014. |
Authors | Hermanrud C, Ryner ML, Engdahl E, Fogdell-Hahn A. |