Aurum™ Total RNA Mini Kit

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Overview

RNA Isolation Selection Guide
  Aurum Total RNA Kits PureZOL RNA Isolation Reagent
Mini Fatty and Fibrous Tissue 96
Format Mini column Filtration (vacuum or spin) Mini column Filtration (vacuum or spin) 96-well plate Filtration (vacuum or spin)
Maximum starting amounts        
   Cultured cells 2 x 106 1 x 106 1 x 107
   Bacterial cells 2.4 x 109 8 x 108 2.4 x 109
   Yeast cells 3 x 107 2 x 107 3 x 107
   Animal tissue — hard 20 mg 100 mg 100 mg
   Animal tissue — soft    to moderately hard 40 mg 100 mg 100 mg
   Plant tissue 40 mg 100 mg 100 mg
Isolation method Silica membrane Lysis with PureZOL reagent, purification on silica membrane Silica membrane Organic extraction
Number of preps 10* or 50 mini preps 10* or 50 mini preps 2 x 96-well plate 50 or 100 preps (1 ml/prep)
Number of washes 3 3 3
DNase I included** Yes Yes Yes No
DNase I digest time 15 min (animal tissue, 25 min) 15 min 10 min
Total preparation time <50min (1.5 hr with DNase I) <50min (1.5 hr with DNase I) <60 min <60 min
Binding capacity >100 µg >100 µg >40 µg
Elution volume 2 x 40 µl 2 x 40 µl 80 µl 30–100 µl
Yield*** 28 µg/2 x 106 NIH 3T3 cells 96 µg/100 mg brain 10 µg/1 x 106 NIH 3T3 cells 1.15 µg/mg brain
  38 µg/2 x 106 HeLa cells 58 µg/100 mg breast 17 µg/1 x 106 HeLa cells 1.37 µg/mg fat
  20 µg/40 mg brain 85 µg/100 mg heart 11 µg/2 x 107 S. cerevisiae cells 1.2 µg/mg heart
  52 µg/20 mg thymus 63 µg/100 mg skin 5 µg/8 x 108 E. coli cells 0.66 µg/mg cartilage
  132 µg/40 mg spleen 54 µg/100 mg cartilage 5 µg/8 x 108 B. cereus cells 1.2 µg/mg skin
  36 µg/40 mg lung     1.0 µg/mg potato
  32 µg/40 mg potato     1.37 µg/1 x 105 293H cells
  28 µg/3 x 107 S. cerevisiae cells     0.1 µg/mg Arabidopsis
  54 µg/2.4 x 109 E. coli cells     0.38 µg/mg A. niger
        0.63 µg/2.4 x 107 E. coli cells
        0.31 µg/3 x 105 S. cerevisiae cells
Purity A260/A280 of 1.9–2.1 A260/A280 of 1.9–2.1 A260/A280 of 1.9–2.1 A260/A280 of 1.9–2.1
 

* Trial sizes of the Aurum total RNA mini kit and Aurum total RNA fatty and fibrous tissue kit are available.

** Removal not required.

*** Yields will vary depending on the development stage, tissue type or cell line, and growth conditions used.

Isolated RNA Quality

Automated electrophoresis of high-quality eukaryotic total RNA. Total RNA was isolated from 30 mg of rat brain using the Aurum total RNA mini kit in the spin format. Start times of the 18S and 28S ribosomal RNAs were 39.6 and 45.5 sec, respectively. The clearly defined peaks and minimal background fluorescence indicate intact, undegraded RNA.

Isolated RNA Performance via qPCR

Real-time RT-PCR with total RNA isolated using the Aurum total RNA mini kit. Total RNA isolated from a culture of 2 x 106 HeLa cells was eluted in Aurum RNA elution solution and a sample used in two-step real-time RT-PCR. Reverse transcription reactions were performed on tenfold dilutions (1 µg to 1 pg) of the HeLa total RNA using the iScript cDNA synthesis kit. cDNA from these reactions was used to detect human β-actin gene expression levels using gene-specific primers and iQ SYBR® Green supermix. Real-time data were acquired on Bio-Rad's iCycler iQ system. A, real-time curves for detection of human β-actin cDNA; B, standard curve showing r = 0.999, efficiency = 103.3%. RFU, realtive fluorescence units.

The Aurum total RNA mini kit produces high-quality DNA-free total RNA from a wide range of starting materials, including cultured cells, bacteria, and yeast, as well as animal and plant tissues. The kit utilizes a stringent reagent composed of guanidine isothiocyanate and β-mercaptoethanol for efficient sample lysis and quick RNase inactivation, followed by purification on silica membrane in a spin-column format using a spin- or vacuum-mediated protocol. The entire purification process, including DNase I digest, is completed in less than 60 minutes. Isolated RNA is of the highest integrity and is suitable for the most discriminating applications. The Aurum total RNA mini kit can also be used for RNA cleanup and desalting.

Features include:

  • High-quality RNA isolation from a wide variety of samples in less than 60 minutes
  • RNase-free reagents and plasticware, including DNase I for removal of genomic DNA, at no additional cost
  • Rapid silica binding-based purification technology in a spin-column format
  • Easy-to-use spin- or vacuum-compatible protocol

Genomic DNA, RNA, and Plasmid Purification Kits
Bio-Rad offers a complete line of products for the isolation and purification of nucleic acids. From the purification of milligram quantities of plasmid DNA for restriction analysis or transformation to the rapid and efficient purification of PCR products, Bio-Rad provides you with high-quality kits that eliminate time-consuming, labor-intensive steps.

Format Mini column
Method Spin- or vacuum-mediated silica binding
Number of preps 50
Maximum starting material amounts 20 mg hard animal tissue
  40 mg plant or soft to moderately hard animal tissues
  2 x 106 cultured cells
  2.4 x 109 bacterial cells
  3 x 107 yeast cells
Binding capacity >100 µg purified total RNA
Purity A260/A280 of 1.9–2.1
RNA size distribution >200 nucleotides
Preparation time 30–60 min (including DNase I digest)
Maximum elution volume 80 µl (2 elutions of 40 µl each)
Minimum elution volume 30 µl*
Yield** 20 µg/40 mg brain
  132 µg/40 mg spleen
  36 µg/40 mg lung
  32 µg/40 mg potato
  54 µg/2.4 x 109 E. coli cells
  28 µg/3 x 107 S. cerevisiae cells
  28 µg/2 x 106 NIH 3T3cells
  38 µg/2 x 109 HeLa cells
*The elution volume is reduced to 30µl
when isolating total RNA from small amounts
of starting material (<500,000 cells
or 10 mg of tissue).
 
**Yields will vary depending on the
developmental stage, tissue type or cell line,
and growth conditions used.
 
Aurum™ Total RNA Mini Kit

732-6820
DNA-free RNA isolation kit, includes RNA binding columns, capless wash and capped microcentrifuge tubes, lyophilized DNase I, RNase-free reagents, for 50 preps

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Number Description Options
6090 Amplification Reagents and Plastics Brochure, Rev D Click to download [ Add to Cart (Free) ]
2920 Aurum Total RNA Mini Kit Product Information Sheet, Rev B Click to download [ Add to Cart (Free) ]
2954 Aurum Total RNA Mini Kit Protocol Overview — Vacuum Format Click to download
2926 Aurum Total RNA Mini Kit Protocol Overview — Spin Format Click to download
4110133 Instruction Manual, Aurum Total RNA Mini Kit, Rev B Click to download
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5990 Gene Expression Analysis Solutions Brochure, Rev C Click to download [ Add to Cart (Free) ]