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

• Very competitive price (cost per test)   
• All reagents stable for > 2 years 
• High purity / standardised enzymes employed 
• Mega-Calc™ software tool is available from our website for hassle-free raw data processing 
• Simple format

The Total Dietary Fiber Assay Kit for the analysis of Total, Soluble and Insoluble Dietary Fiber according to AOAC and AACC approved methods.

Introduction

Discover our Yeast Extract Peptone Dextrose (YPD) Agar, designed for the cultivation and growth of yeasts and fungi, including Saccharomyces cerevisiae. This nutrient-rich medium supports robust microbial proliferation, making it ideal for molecular biology, fermentation, and genetic research.

Discover our Yeast Extract Peptone Dextrose (YPD) Agar, designed for the cultivation and growth of yeasts and fungi, including Saccharomyces cerevisiae. This nutrient-rich medium supports robust mi……

cfDNA Purification Kit (Magnetic Beads)

The cfDNA Purification Kit (Magnetic Beads) was developed for cell free DNA (cfDNA) enrichment by separating genomic DNA contamination from isolated cfDNA samples.

Many diagnostic technologies for detection of disease signals in cfDNA begin with isolation and purification of DNA from liquid biopsy that include urine, plasma, cerebrospinal fluid. The most widely explored biotechnology is assays used to detect cancer-derived plasma cfDNA. Silica-based magnetic bead cfDNA isolation kits can reliably extract total DNA from plasma, but typically yield a large variation in cfDNA that includes the presence of genomic DNA that often depends on tumor stage, tumor size, or healthy status individuals. Most of the commercial cfDNA isolation kits can’t specifically recover the cfDNA while leaving the high molecular weight genomic DNA behind. The presence of genomic DNA can lead to decreased sensitivity or inconsistent results in downstream applications such as next-generation sequencing (NGS), PCR, QPCR, and digital PCR etc.

Therefore, an additional purification step to enrich cfDNA before downstream methods helps to improve signal from fragments that originate from cancer cells. A proportion of cancer-derived cfDNA fragment signals are below 100 bp and are often not detectable except by qPCR or single-stranded DNA based library preparation for NGS (1, 2, 3). Furthermore only 1% of cancer-derived fragments are found above 400 bp (1, 2). Capture of size-selected fragments between 90-150 bp improved detection of cancer by 2-4 fold (4). Furthermore, TF-bound and protected cfDNA fragments are also being investigated for active cancer-specific signals down to 35-80 bp (5, 6).

This kit uses Dual Solid Phase Reversible Immobilization (SPRI) technology for cfDNA purification. Most Dual SPRI procedures do NOT recover fragments below 100 bp. The kit can be used for the enrichment of cfDNA isolated from liquid biopsies, plasma, serum, and urine. The kit separates cfDNA (50-500 bp) and genomic DNA, and recovers of 90% of the cfDNA without the high molecular weight genomic DNA with high efficiency. Fragments at 500 bp and above may also be retained. Both the 50-500 bp and >500 bp DNA fractions can be used for downstream applications such as single-stranded or double stranded NGS library prep, qPCR, ddPCR, and other methods.

Features

• Separation of cfDNA and genomic DNA; Recovery of both types of DNA
• Recovery of cfDNA (50-500 bp)
  • As short as 50 bp can be recovered
• Recovery of high molecular weight genomic DNA
• Removal of unwanted components and other impurities
• Automation friendly

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Examples of cfDNA purification. Both cfDNA and genomic DNA can be recovered separately.

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The range of recovered small DNA fragments is from 50 to 500 bp. The input DNA are mixtures of sheared small DNA fragments and intact genomic DNA. The ratios of sheared DNA fragments versus genomic DNA are indicated.

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Recovery rates of cfDNA and genomic DNA.

Many diagnostic technologies for detection of disease signals in cfDNA begin with isolation and purification of DNA from liquid biopsy that include urine, plasma, cerebrospinal fluid. The most widely explored biotechnology is assays used to detect cancer-derived plasma cfDNA. Silica-based magnetic bead cfDNA isolation kits can reliably extract total DNA from plasma, but typically yield a large variation in cfDNA that includes the presence of genomic DNA that often depends on tumor stage, tumor size, or healthy status individuals. Most of the commercial cfDNA isolation kits can’t specifically recover the cfDNA while leaving the high molecular weight genomic DNA behind. The presence of genomic DNA can lead to decreased sensitivity or inconsistent results in downstream applications such as next-generation sequencing (NGS), PCR, QPCR, and digital PCR etc.