Total RNA Purification Maxi Kit

Product Description

• Isolate total RNA, from large rRNA down to microRNA (miRNA)
• For mammalian cells and tissue, blood, bacteria, yeast, fungi, plants, and more
• Isolate high quality total RNA with complete size range from a variety of sources
• No phenol or chloroform extractions
• Rapid spin column format, protocol completed in 40min
• Kits for smaller input amounts and 96-well kit for high throughput applications also available

For large serum and plasma input amounts (1 ml - 5ml) please use the Plasma/Serum Circulating RNA Purification Kit (see link below).

Norgen's Total RNA Purification Maxi Kit provides a rapid method for the isolation and purification of total RNA from large amounts of cultured animal cells, tissue samples, blood, bacteria, yeast and fungi. 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. The purified RNA is of the highest integrity, 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.

Purification Technology

Purification is based on spin column chromatography using Norgen's silicon carbide (SiC) resin as the separation matrix.

The process involves first lysing the cells or tissue of interest with the provided Lysis Solution. Binding solution and ethanol are then added to the lysate, and the solution is loaded onto a 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 total RNA is eluted with the elution buffer. The purified RNA is of the highest integrity, and the complete profile of RNA species is represented.

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.

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 uL reverse transcription using Invitrogen Superscript III. Three microliters of the reverse transcription was used as the template in a 20 uL 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.

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 for discovery
• microRNA cloning and amplification
• PCR-based virus detection
• PCR-based viable bacteria detection
• Northern Blotting
• RNase Protection
• Primer Extension