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Total RNA Clean-Up and Concentration Kit Add to Cart

Cat#:	43200-NB
Quantity:	100 preps
Price:	542 €
Supplier:	Norgen
Shipping:	RT

User Manual

For rapid and efficient clean-up and concentration of total RNA, including microRNA, without phenol

Please note: This kit can be used instead of discontinued #52900-NB

• Clean-up and concentrate total RNA isolated by different methods including phenol-chloroform extractions or alcohol precipitations
• Fast and easy processing - Rapid spin-column format allows for the processing of 10 samples in 20 minutes, while the 96-well plates can be processed in 30 minutes
• No phenol or chloroform extractions
• Clean-up RNA from contaminants including enzymes, RNases and nucleotides
• Isolate all small RNA molecules including microRNA, siRNA, tRNA, and 5S rRNA
• Purified RNA is fully compatible with all downstream applications
• Also available in a 96-well format for high throughput applications

Norgen’s RNA Clean-Up and Concentration Micro Kit provides a rapid method for the purification, cleanup and concentration of up to 35 µg of RNA isolated using different methods including phenol/guanidine-based protocols, and from various upstream enzymatic reactions such as DNase treatment, labeling and in vitro transcription. 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 reaction components such as proteins, RNases and nucleotides, 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 end-point or quantitative reverse transcription PCR, Northern blotting, RNase protection and primer extension, and expression array assays.

Workflow

Effective Clean-Up to Produce High Quality Total RNA with Complete Size Diversity. Norgen’s RNA Clean-Up and Concentration Kit effectively cleans up RNA isolated from phenol-based extractions without the loss of RNA diversity by retaining all RNA species including small RNAs. Total RNA was isolated from 5 x 108 E. coli using a competitor’s phenol-based RNA extraction reagent. The resulting RNA was then purified using Norgen’s RNA Clean-Up and Concentration Kit. As controls, total RNA was extracted using both Norgen´s Total RNA Purfication Kit (#17200, no phenol required) and the phenol-based RNA extraction reagent only without any clean-up. Resolution of 7 µL of the 50 µL purified RNA on a 1X MOPS, 1.5% formaldehyde-agarose gel showed the RNA extracted with phenol was successfully cleaned-up by Norgen’s RNA Clean-Up and Concentration Kit without the loss of small RNA species.

Effective Clean-Up to Produce High Quality Total RNA Compatible to Bioanalyzer Analysis. Total RNA was isolated from 0.75 million HeLa cells using a competitor’s phenol-based RNA extraction reagent. The resulting RNA was then purified using Norgen’s RNA Clean-Up and Concentration Kit. As controls, total RNA was extracted using both Norgen´s Total RNA Purfication Kit (#17200, no phenol required) or the phenol-based RNA extraction reagent only without clean-up. Resolution of 1 µL of the 50 µL purified RNA on an Agilent RNA Nano 6000 Chip showed that without clean-up, the RNA sample isolated using the phenol-based RNA extraction reagent only resolved poorly on a bioanalyzer. In contrast, RNA extracted with the phenol-based RNA extraction reagent followed by clean-up by Norgen’s RNA Clean-Up and Concentration Kit resolved properly on the bioanalyzer showing excellent quality.

Effective Concentration and Detection of Low Amounts of RNA Input. Norgen’s RNA Clean-Up and Concentration Kit can effectively concentration RNA inputs in the picogram range. Increasing amounts of HeLa RNA in 50 µL input volumes were concentrated to 20 µL using Norgen’s RNA Clean-Up and Concentration Kit. A 10 µL aliquot of the purified RNA was then used as the template in a qRT-PCR reaction to detect the S14 gene. The amounts indicated on the graph correspond to the amount of RNA that was used as the input for the qRT-PCR reaction, demonstrating the consistent performance of the kit even in the picogram range.