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RNA/DNA/Protein Purification Plus Kit

For sequential isolation of total RNA, genomic DNA and total proteins from the same sample

Norgen's RNA/DNA/Protein Purification Plus Kit provides a rapid method for the isolation and purification of total RNA, genomic DNA and proteins sequentially from a single sample of cultured animal cells, small tissue samples, blood, bacteria, yeast, fungi or plants. The total RNA, genomic DNA and proteins are all column purified in less than 30 minutes (Application 1). This kit is ideal for researchers who are interested in studying the genome, proteome and transcriptome of a single sample, such as for studies of microRNA profiling, gene expression including gene silencing experiments or mRNA knockdowns, studies involving biomarker discovery, and for characterization of cultured cell lines. Norgen's RNA/DNA/Protein Purification Plus Kit is especially useful for researchers who are isolating macromolecules from precious, difficult to obtain or small samples such as biopsy materials or single foci from cell cultures, as it eliminates the need to fractionate the sample. Furthermore, analysis will be more reliable since the RNA, DNA and proteins are derived from the same sample, thereby eliminating inconsistent results. The purified macromolecules are of the highest purity and can be used in a number of different downstream applications.

Features

  • Complete column purification - The RNA, DNA and proteins are all column purified
  • Reduce variability - RNA, DNA and proteins are isolated from a single sample with no splitting of the lysate, thus reducing inconsistent results and variability
  • Isolate from small samples - Sequential isolation of RNA, DNA and protein from a single sample. Ideal for precious, difficult to obtain or small samples such as biopsy material or single foci from cell cultures.
  • Rapid procedure - Isolate total RNA, genomic DNA and total proteins from a single sample in less than 30 minutes
  • Isolate a diversity of RNA species - All sizes of RNA are isolated, from large mRNA down to microRNA
  • Isolate high quality RNA, DNA and proteins - The purified RNA, DNA and proteins are of the highest quality and can be used in a number of downstream applications

Application 1 : Recovery of True Total RNA including microRNA from HEK-293 Cells.

Panel A is a 1X MOPS 1% agarose gel showing the RNA that was isolated from 2 different samples of ~ 800,000 HEK-293 cells using either Norgen's RNA/DNA/Protein Purification Plus Kit or a competitor's multiple-analyte purification kit. Both kits isolated large RNA (represented by 28S and 18S rRNA) with high integrity but Norgen's RNA/DNA/Protein Purification Plus Kit provided the added benefit of recovering small RNA without any additional protocol. Panel B is a result from a bioanalyzer resolution of the eluted RNA. Similar to the agarose gel, Norgen's RNA/DNA/Protein Purification Plus Kit showed the added benefit of recovering small RNA. The difference in small RNA recovery was also demonstrated by gene-specific RT-qPCR. One microgram of RNA was used in RT-qPCR reactions for human S15 (for Large RNA) and miR-21 (for microRNA) genes. The RNA isolated by Norgen's RNA/DNA/Protein Purification Plus Kit showed similar Ct value to RNA isolated by the competitor's kit for the large RNA (Panel D). However, Norgen's RNA/DNA/Protein Purification Plus Kit showed superior recovery of small RNA including microRNAs as depicted by the miR-21 RT-qPCR (Panel C).

Application 2 : Recovery of True Total RNA including microRNA from Hamster Liver.

Panel A is a 1X MOPS 1% agarose gel showing the RNA that was isolated from 2 different samples of 15 mg hamster liver using either Norgen's RNA/DNA/Protein Purification Plus Kit or a competitor's multiple-analyte purification kit. Both kits isolated large RNA (represented by 28S and 18S rRNA) with high integrity but Norgen's RNA/DNA/Protein Purification Plus Kit provided the added benefit of recovering small RNA without any additional protocol. Panel B is a result from a bioanalyzer resolution of the eluted RNA. Similar to the agarose gel, Norgen's RNA/DNA/Protein Purification Plus Kit showed the added benefit of recovering small RNA. The difference in small RNA recovery was also demonstrated by gene-specific RT-qPCR. One microgram of RNA was used in RT-qPCR reactions for hamster beta-Actin (for Large RNA) and miR-21 (for microRNA) genes. The RNA isolated by Norgen's RNA/DNA/Protein Purification Plus Kit showed similar Ct value to RNA isolated by the competitor's kit for the large RNA (Panel D). However, Norgen's RNA/DNA/Protein Purification Plus Kit showed superior recovery of small RNA including microRNAs as depicted by the miR-21 RT-qPCR (Panel C).

Application 3 : Recovery of Intact, High Quality Genomic DNA from HEK-293 cells and Hamster Liver.

Panel A is a 1% agarose gel showing the gDNA isolated from the same HEK-293 cell or hamster liver samples using Norgen's RNA/DNA/Protein Purification Plus Kit or competitor's multiple-analyte purification kit. Lane M is Norgen's HighRanger 1 kb DNA Ladder and the sample lanes contain 10 µL of each of the 100 µL elutions. The gel showed high quality, and intact genomic DNA, with a better yield using Norgen's RNA/DNA/Protein Purification Plus Kit. Panel B is the result of qPCR amplification of 25 ng of eluted genomic DNA using 5S rRNA-specific primers. Genomic DNA isolated using Norgen's RNA/DNA/Protein Purification Plus Kit is of high quality and performed similar to or better than competitor's product.

Application 4 : High Quality Total Proteins Eluted in Mass Spec-Compatible Buffer.

Norgen's RNA/DNA/Protein Purification Plus Kit provides an additional column purification step for effective concentration and clean-up of the isolated proteins. In contrast, most competing multiple analyte isolation products require protein precipitation and the precipitated proteins are required to be resuspended in buffer with high-detergent content (such as SDS-PAGE loading dye) for full recovery. Protein fractions (from hamster liver) isolated by Norgen's RNA/DNA/Protein Purification Plus Kit and a competitor's kit were resolved on a 12% SDS-PAGE protein gel. Panel A showed that when the competitor's precipitated protein fraction was resuspended in a provided SDS-PAGE loading buffer, the protein recovery was similar among the two kits. Panel B showed that when the same precipitated protein fraction from the competitor's kit was resuspended in a Tris-based buffer containing no detergent or denaturant, the protein recovery became drastically reduced. In contrary, Norgen's RNA/DNA/Protein Purification Plus Kit purified proteins by column and the eluted proteins are already in a buffer compatible with most downstream applications including mass spectrophotometry as well as standard protein quantification methods (including Bradford assays).

Kit Specificaations

Maximum Column Binding Capacity 50 µg for RNA, 20 µg for DNA, 200 µg for protein
Maximum Column Loading Volume 650 µL
Size of RNA Purified All sizes, including small RNA (<200 nt)
Size of DNA Purified ≥ 30 kb
Maximum Amount of Starting Material Animal Cells: 5 x 106 cells
Animal Tissues: 25 mg (for selected tissues)
Blood: 100 µL
Bacteria: 1 x 109 cells
Yeast: 1 x 108 cells
Fungi, Plant : 50 mg
Time to Complete 10 Purifications 30 minutes
Average Yields HeLa Cells (1 x 106 cells): 15 µg RNA, 8 µg DNA, 150 µg protein

Protocol

RNA/DNA/Protein Purification Plus Kit - Protocol (PDF)
RNA/DNA/Protein Purification Plus Kit - Product Information (PDF)

Ordering Information

Product Cat.No. Storage Quantity Price(US$)  
RNA/DNA/Protein Purification Plus Kit 47700 Room Temp. 50 preps 535.00 Buy