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Norgen Biotek Corp. is dedicated to providing our customers with first class sample preparation kits for RNA, microRNA, DNA and protein purification, clean-up and concentration and to providing dedicated and expert support services to our customers and partners worldwide.
As a testament of our commitment to quality, Norgen is proud to be an ISO 9001:2008 and ISO 13485:2003 registered company.
It is a pleasure to serve you and we look forward to working closely with you to provide the best sample preparation products for your specific needs. |
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Please contact us at:
info@norgenbiotek.com
Norgen Biotek Corp.
3430 Schmon Parkway,
Thorold
Ontario, Canada, L2V 4Y6
Tel: (905) 227-8848
Fax: (905) 227-1061 |
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Total RNA Purification Maxi Kit
(Cat# 26800)
- Isolate true total RNA including siRNA and microRNA
- No phenol extractions
- Purify high-quality RNA in 40 minutes
- Isolate total RNA from a wide variety of sources
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Norgen’s Total RNA Purification Maxi Kit provides a rapid method for the isolation and purification of total RNA from cultured animal cells, tissue samples, blood, bacteria, yeast, fungi, plants and viruses. The kit purifies all sizes of RNA, from large mRNA and ribosomal RNA down to microRNA (miRNA) and small interfering RNA (siRNA), without the use of phenol or chloroform. Norgen's kit is the only kit on the market that isolates true total RNA, as other kits must use phenol to recover all sizes of RNA. Therefore Norgen's kit offers significant advantages in functionality, savings on cost, ease-of-use, no hazardous organic waste, and no inhibitory effect on PCR amplifications as a result of residual phenol. With this kit both miRNA and mRNA can be studied from the same sample without further purifications, thus offering considerable advantages when comparing and relating expression of miRNA to other RNA.
Purification is based on spin column chromatography using Norgen’s proprietary resin as the separation matrix. Briefly, the cells or tissue of interest are first lysed using the provided Lysis Solution, ethanol is added and the RNA is bound to Norgen's maxi column (BIND). Under these conditions only the RNA will bind to Norgen's resin while most of the contaminating cellular proteins are removed in the flowthrough or retained on top of the resin. The bound RNA is then washed to remove any remaining impurities (WASH). Lastly, the purified total RNA is eluted into 2 mL of the provided Elution Buffer or water (ELUTE). Please see procedure flowchart to the right.
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 FEATURES AND BENEFITS
- Isolate a diversity of RNA species - All RNA species can be isolated, from large mRNA and ribosomal RNA down to microRNA (miRNA) and small interfering RNA (siRNA)
- Fast and easy processing - Rapid spin-column format allows for the processing of 4 samples in under 40 minutes.
- No phenol:chloroform extractions – Total RNA is isolated without the use of harmful chemicals such as phenol or chloroform.
- Isolate total RNA from a wide variety of sources – Total RNA can be isolated from cultured animal cells, tissue samples, blood, bacteria, yeast, fungi, plants and viruses using this kit.
- Isolate inhibitor-free RNA 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
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APPLICATIONS
- Quantitative, real-time RT-PCR for both large mRNA and small RNA including miRNA
- RT-PCR for both large mRNA and small RNA including miRNA
- Expression profiling
- Next Generation Sequencing for RNA and miRNA
- miRNA from plasma for discovery
- microRNA cloning and amplification
- PCR-based virus detection
- PCR-based viable bacteria detection
- Northern blotting
- RNase protection
- Primer extension
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Figure 1. Isolate a Diversity of RNA Species. Total RNA was isolated from 10 mL of an overnight E. coli culture using Norgen’s Total RNA Purification Maxi Kit. One microliter of the 2 mL elution was analyzed on an Agilent® 2100 BioAnalyzer RNA Nano 6000 chip. As it can be seen, high quality total RNA was isolated with minimal degradation. As well, a large amount of small RNA (<200 nt) was recovered, indicating that the kit is able to isolate a diversity of RNA species.
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Figure 2. Good Amplification and Consistency of Isolation. Norgen’s Total RNA Purification Maxi Kit was used to isolate total RNA from six different overnight samples of E. coli (10 mL samples). In brief, one microgram of the isolated RNA was subjected to a 20 µL reverse transcription using Invitrogen Superscript III. Three microliters of the reverse transcription was used as the template in a 20 µL qPCR reaction using16S rRNA E. coli specific primers and Bio-Rad iQ mix. As it can be seen, the RNA was detected and successfully amplified in all cases. Furthermore, all samples showed a similar CT value, indicating consistent isolation. |
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Kit Specifications
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Column Binding Capacity
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1.5 mg
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Maximum Column Loading Volume
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20 mL
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Size of RNA Purified
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All sizes, including small RNA (<200 nt)
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Maximum Amount of Starting Material:
Animal Cells
Animal Tissues
Blood
Bacteria
Yeast
Fungi
Plant Tissues
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5 x 107 cells
50 - 250 mg
2 – 10 mL*
2.5 x 1010 cells
2 x 108 cells
1 g
1 g
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Time to Complete 4 Purifications
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40 minutes
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Average Yields
HeLa Cells (5 x 107 cells)
E. coli (2.5 x 1010 cells)
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750 μg
1.5 mg
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* for isolating total RNA from purified leukocytes
Kit Contents
1. Lysis Solution
2. Wash Solution
3. Elution Solution
4. RNA Maxi Spin Columns (with collection tubes)
5. Elution tubes (50 mL)
6. Product Insert
7. Lysis Solution
8. Wash Solution
Storage Conditions and Product Stability
All solutions should be kept tightly sealed and stored at room temperature. These reagents should remain stable for at least 1 year in their unopened containers. |
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| PROTOCOL |
Total RNA Purification Maxi Kit - Protocol
PDF version
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| PRODUCT INFORMATION SHEETS |
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| APPLICATION NOTES |
Total RNA Purification Maxi Kit - Product Information Sheet
PDF version
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POSTERS |
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| MSDS |
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| SUPPLEMENTARY PROTOCOLS |
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| REFERENCES |
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| FREQUENTLY ASKED QUESTIONS |
Total RNA Purification Maxi Kit - Frequently Asked Questions
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Order NOW at our online store:
Item |
Cat. No. |
Kit Size |
Buy Online |
Total RNA Purification Maxi Kit |
26800 |
8 preps |
Buy Now |
Order by Phone: Toll Free (USA & CANADA): 1-866-NORGENB (667-4362) or from anywhere at (905)227-8848.
Order by Email: order by sending an email to orders@norgenbiotek.com.
Order by Fax: Fax your order to (905)227-1061
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