These kits provide a fast, reliable and convenient method to purify and concentrate high quality, high purity and inhibitor-free cell-free circulating and exosomal RNA using a convenient spin column method. These kits can purify RNA from fresh or frozen serum or plasma samples prepared from blood collected on either EDTA or Citrate. Plasma samples prepared from blood collected on heparin should not be used, as heparin can significantly interfere with many downstream applications such as RT-PCR. The purified plasma/serum RNA is fully compatible with all downstream applications including PCR, qPCR, methylation-sensitive reverse transcription qPCR, reverse transcription PCR, Northern blotting, RNase protection and primer extension, expression array assays, and NGS.
Background
Plasma/Serum cell-free circulating RNA or exosomal RNA has the potential to provide biomarkers for certain cancers and disease states. Exosomes are 40 – 150 nm membrane vesicles, which are secreted by most cell types. Exosomes can be found in saliva, blood, urine, amniotic fluid and malignant ascitic fluids, among other biological fluids. Evidence has been accumulating recently that these vesicles act as cellular messengers, conveying information to distant cells and tissues within the body. These exosomes may play a functional role in mediating adaptive immune responses to infectious agents and tumours, tissue repair, neural communication and transfer of pathogenic proteins. For this reason exosomal RNAs may serve as biomarkers for various diseases including cancer. As the RNA molecules encapsulated within exosomes are protected from degradation by RNAses they can be efficiently recovered from biological fluids, such as plasma or serum.
Plasma/Serum RNA Purification Mini Kit
This kit can purify RNA from fresh or frozen serum or plasma samples prepared from blood collected on either EDTA or Citrate, from volumes ranging from 50 µL to 200 µL. The purified plasma/serum RNA is eluted in a flexible final volume of 10 µL to 25 µL.
Plasma/Serum RNA Purification Midi Kit
This utilizes a two column method, and can purify RNA from fresh or frozen serum or plasma samples prepared from blood collected on either EDTA or Citrate, from volumes ranging from 250 µL to 1.5 mL. The first column will handle the large volume input of bodily fluids that is followed by a concentration on a mini column for a final elution of 50 µL to 100 µL.
Plasma/Serum RNA Purification Maxi Kit
This kit can purify RNA from fresh or frozen serum or plasma samples prepared from blood collected on either EDTA or Citrate, from volumes ranging from 2 mL to 5 mL. The first column will handle the large volume input of bodily fluids that is followed by a concentration on a mini column for a final elution of 50 µL to 100 µL.
Isolate RNA after Purifying EVs and Exosomes
For Ultracentrifugation, Exoquick, and Filtration
| Cat. # | Name | Elution Volume | Plasma/Serum | Urine | Cell-Culture Media |
|---|---|---|---|---|---|
| 55000 | Plasma/Serum RNA Purification Mini Kit | 10 – 25 µL | 50 µL – 1 mL | 250 µL – 1 mL | 5 – 10 mL |
| 35300 | Total RNA Purification Micro Kit | 20 – 50 µL | 1 – 4 mL | 2 – 10 mL | 10 – 20 mL |
| 17200 | Total RNA Purification Kit | 50 – 100 µL | 4 – 10 mL | 11 – 30 mL | 20 – 35 mL |
Figure 1 / 13
Click for expanded view
| Kit Specifications | |
| Minimum Plasma/Serum Input | 2 mL |
| Maximum Plasma/Serum Input | 5 mL |
| Size of DNA Purified | All sizes, including miRNA and small RNA (<200 nt) |
| Elution Volume | 50-100 μL |
| Time to Complete 10 Purifications | 35-40 minutes |
| Average Yields* | Variable depending on specimen |
*Please check page 7 of the Product Insert for Average Plasma/Serum Yields and Common RNA Quantification Methods.
This kit is suitable for the isolation of RNA from fresh or frozen serum or plasma prepared from blood collected on either EDTA or Citrate. Plasma samples prepared from blood collected on heparin should not be used as heparin can significantly interfere with many downstream applications such as RT-PCR.
Storage Conditions and Product Stability
All solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 2 years after the date of shipment. It is recommended to warm Lysis Buffer A for 20 minutes at 60°C if any salt precipitation is observed.
| Kit Components | Cat. 55000 (50 preps) | Cat. 56100 (20 preps) | Cat. 56200 (10 preps) |
|---|---|---|---|
| Lysis Buffer A | 2 x 20 mL | 100 mL | 1 x 130 mL 1 x 30 mL |
| Wash Solution A | 18 mL | 1 x 38 mL 1 x 18 mL | 38 mL |
| Elution Solution A | 6 mL | 6 mL | 6 mL |
| Elution Buffer F | – | 15 mL | 15 mL |
| Micro Spin Columns | 50 | – | – |
| Mini Spin Columns | – | 20 | 10 |
| Midi Spin Columns | – | 20 | – |
| Maxi Spin Columns | – | – | 10 |
| Collection Tubes | 50 | 20 | 10 |
| Elution Tubes (1.7 mL) | 50 | 20 | 10 |
| Product Insert | 1 | 1 | 1 |
These kits are designed for the rapid preparation of plasmid DNA from Escherichia coli.
Plasmid MiniPrep Kit
This kit is designed for the rapid preparation of plasmid DNA from small cultures of Escherichia coli using convenient spin columns. The plasmid DNA is preferentially purified from other cellular components such as genomic DNA and RNA. This kit is able to purify plasmids up to 13,000 bp in size, and the typical purification yield is up to 20 μg from 1.5 mL of bacterial culture. Purified DNA is of excellent quality for transformation, restriction enzyme digestion, sequencing and more. Also available in a 96-well format.
Plasmid MiniPrep Kit (Magnetic Bead System and High Throughput Magnetic Bead System)
Norgen’s Plasmid MiniPrep Kit (Magnetic Bead System) is designed for the rapid preparation of plasmid DNA from small batch cultures of Escherichia coli. Norgen’s magnetic beads bind DNA under optimized salt concentrations and release the bound DNA under low salt and slightly alkali conditions. The plasmid DNA is preferentially purified from other cellular components such as genomic DNA and RNA. The purified plasmids are fully digestible with all restriction enzymes tested, and are completely compatible with real-time PCR and NGS.
Norgen’s Plasmid MiniPrep Kit (Magnetic Bead System) is also available in a 96-well (HT) format for high throughput applications. Purification with the 96-well plates can be integrated with a robotic automation system.
Plasmid MaxiPrep Kit
This kit is designed for the rapid spin column preparation of plasmid DNA from up to 100 mL of Escherichia coli cultures. The kit allows for the isolation of plasmid DNA with final endotoxin levels of 0.1 EU/µg of DNA or less. The kit is able to purify plasmids up to 13,000 bp in size, and typical yields from a 100 mL culture for a high copy number plasmid are between 0.4 and 1.0 mg. The purified DNA is fully digestible with all restriction enzymes tested, and is completely compatible with manual or automated sequencing to achieve 95-100% accuracy.
Figure 1 / 5
Click for expanded view
| Kit Specifications | |
| Column Binding Capacity | 25 µg |
| Size of Plasmids Purified | Up to 13,000 bp |
| Average Yield from 1.5 mL of Culture | Up to 20 µg |
| Time to Complete 10 Purifications | 30 minutes |
Storage Conditions and Product Stability
All solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 1 year after the of shipment. The RNase should be stored at -20°C upon arrival. The Resuspension Solution AZ should be stored at 4oC upon addition of RNase enzyme.
| Component | Cat. 13300 (50 preps) | Cat. 46400 (250 preps) | Cat. 46500 (4 preps) | Cat. 46600 (20 preps) | Cat. 60300 (50 preps) | Cat. 63000 (192 preps) |
|---|---|---|---|---|---|---|
| Resuspension Solution AZ | 12 mL | 60 mL | 20 mL | 100 mL | 12 mL | 2 x 20 mL |
| Lysis Buffer N | 40 mL | 80 mL | 40 mL | 2 x 80 mL | 40 mL | 2 x 40 mL |
| Buffer TN | 20 mL | 130 mL | 55 mL | 2 x 130 mL | 20 mL | 1 x 55 mL 1 x 20 mL |
| Wash Solution E | 12 mL | 2 x 18 mL | – | – | – | – |
| Elution Buffer K | 8 mL | 30 mL | – | – | 8 mL | 2 x 8 mL |
| Wash Solution J | – | – | 25 mL | 3 x 25 mL | – | – |
| Elution Buffer J | – | – | 24 mL | 120 mL | – | – |
| RNase A | 1 vial | 1 vial | 1 vial | 1 vial | 1 vial | 1 vial |
| Magnetic Bead Suspension | – | – | – | – | 1 x 1.1 mL | 4 x 1.1 mL |
| Spin Columns | 50 | 250 | – | – | – | – |
| Collection Tubes | 50 | 250 | – | – | – | – |
| DNA Maxi Spin Columns with Collection Tubes (Clear ring in column) | – | – | 4 | 20 | – | – |
| Maxi Spin Filter Columns with Collection Tubes (Grey ring in column) | – | – | 4 | 20 | – | – |
| 96-Well Plate | – | – | – | – | – | 2 |
| Elution Tubes (1.7 mL) | 50 | 250 | – | – | 50 | – |
| Elution Tubes (50 mL) | – | – | 4 | 20 | ||
| 96-Well Elution Plate | – | – | – | – | – | 2 |
| Adhesive Tape | – | – | – | – | – | 2 |
| Product Insert | 1 | 1 | 1 | 1 | 1 | 1 |
K-TDFR-200A
SKU: 700004347
| Content: | 100 assays / 200 assays |
| Shipping Temperature: | Ambient |
| Storage Temperature: | Short term stability: 2-8oC, Long term stability: See individual component labels |
| Stability: | > 2 years under recommended storage conditions |
| Analyte: | Dietary Fiber |
| Assay Format: | Enzymatic |
| Detection Method: | Gravimetric |
| Signal Response: | Increase |
| Limit of Detection: | 0.5 g/100 g |
| Total Assay Time: | ~ 100 min |
| Application examples: | Food ingredients, food products and other materials. |
| Method recognition: | AACC Method 32-05.01, AACC Method 32-06.01, AACC Method 32-07.01, AACC Method 32-21.01, AOAC Method 985.29, AOAC Method 991.42, AOAC Method 991.43, AOAC Method 993.19, CODEX Method Type I and GB Standard 5009.88-2014 |
The Total Dietary Fiber Assay Kit for the analysis of Total, Soluble and Insoluble Dietary Fiber according to AOAC and AACC approved methods.
View Dietary Fiber Measurement Guide – Which Method for which sample?
Dietary fiber can generally be described as the carbohydrate content of food that is not digested in the human small intestine. It passes into the large intestine where it is partially or fully fermented. These characteristics of dietary fiber are associated with its numerous well documented health benefits.
Dietary Fiber is a mixture of complex organic substances, including hydrophilic compounds, such as soluble and insoluble polysaccharides and non-digestable oligosaccharides, as well as a range of non-swellable, more or less hydrophobic, compounds such as cutins, suberins and lignins. The procedures for the determination and analysis of total dietary fiber as outlined in our assay protocol are based on the methods of Lee et al.1 and Prosky et al.2,3 (AOAC 991.43, AOAC 985.29, AACC 32-07.01 and AACC 32-05.01). However, the enzymes in the Megazyme Total Dietary Fiber Kit can also be used in other dietary fiber analytical methods such as AACC Method 32-21.01 and AACC Method 32-06.01.
1. Association of Official Analytical Chemists. (1985). Official Methods of Analysis, 14th ed., 1st suppl. Secs. 43, A14-43, A20, p.399.
2. Association of Official Analytical Chemists. (1986). Changes in methods. J. Assoc. Off. Anal. Chem., 69, 370.
3. Association of Official Analytical Chemists. (1987). Changes in methods. J. Assoc. Off. Anal. Chem., 70, 393.
See General Referee Reports: Journal of AOAC INTERNATIONAL, Vol. 81, No. 1, 1998.
Two separate methods are described in the associated assay protocol:
METHOD 1: DETERMINATION OF TOTAL, SOLUBLE AND INSOLUBLE DIETARY FIBER
Based on AOAC Method 991.43 “Total, Soluble, and Insoluble Dietary Fiber in Foods” (First Action 1991) and AACC Method 32-07.01 “Determination of Soluble, Insoluble, and Total Dietary Fiber in Foods and Food Products” (Final Approval 10-16-91).
METHOD 2: DETERMINATION OF TOTAL DIETARY FIBER
Based on AACC method 32-05.01 and AOAC Method 985.29.
Note that a letter of endorsement from the original method developer, Dr. Leon Prosky, is included in the Documents Tab.
See our full range of starch and dietary fiber products.
Validation of Methods
Advantages
The Total Dietary Fiber Assay Kit for the analysis of Total, Soluble and Insoluble Dietary Fiber according to AOAC and AACC approved methods.