K-AVCHO

SKU: 700004267

100 Assays of each per kit

Content:	100 assays of each per kit
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:	Available Carbohydrates, Dietary Fiber
Assay Format:	Spectrophotometer
Detection Method:	Absorbance
Wavelength (nm):	340
Signal Response:	Increase
Linear Range:	4 to 80 μg of D-glucose, D-fructose or D-galactose per assay
Limit of Detection:	1.475 g/100 g
Reaction Time (min):	~ 5 h
Application examples:	Food ingredients, food products and other materials.
Method recognition:	AOAC Method 2020.07

The Available Carbohydrates Assay Kit method is suitable for the determination of available carbohydrates (AVCHO) comprising *total digestible starch (TDS) plus maltodextrins, sucrose, D-glucose, D-fructose and lactose. New Improved method receiving ‘First Action’ status: AOAC 2020.07. This method is designed to simulate in vivo conditions in the human small intestine (i.e. a 4 h incubation time with PAA + AMG) in parallel with recent advances in Dietary Fiber (DF) methodology (K-RINTDF: AOAC Method 2017.16) and in accordance with the new (physiological based) definition of DF announced by Codex Alimentarius in 2009. Also, sucrose is hydrolysed with a specific “sucrase” enzyme which (unlike invertase which has been used traditionally for this reaction) has no action on fructo-oligosaccharides (FOS).

* Total digestible starch (TDS) is defined as starch that is digested in a 4 h period and is part of the carbohydrate that is available for digestion and absorption in the human small intestine.

See our full range of dietary fiber assay kits.

The Available Carbohydrates Assay Kit method is suitable for the determination of available carbohydrates (AVCHO) comprising *total digestible starch (TDS) plus maltodextrins, sucrose, D-glucose, D-fructose and lactose. New Improved method receiving ‘First Action’ status: AOAC 2020.07. This method is designed to simulate in vivo conditions in the human small intestine (i.e. a 4 h incubation time with PAA + AMG) in parallel with recent advances in Dietary Fiber (DF) methodology (K-RINTDF: AOAC Method 2017.16) and in accordance with the new (physiological based) definition of DF announced by Codex Alimentarius in 2009. Also, sucrose is hydrolysed with a specific “sucrase” enzyme which (unlike invertase which has been used traditionally for this reaction) has no action on fructo-oligosaccharides (FOS).

Introduction

The HiPure Total RNA Kit provides fast purification of high-quality RNA from cells, tissues, and yeast using silica-membrane spin columns with a binding capacity of 100μg RNA. There is no need for phenol/chloroform extractions and time-consuming steps such as CsCl gradient ultracentrifugation or isopropanol precipitation. RNA purified using the HiPure Total RNA Purification System can be used for applications such as RT-PCR, Northern blotting, poly A+ RNA (mRNA) purification, nuclease protection, and in vitro translation.

Details

Specifications

Features	Specifications
Main Functions	Isolation total RNA (not include miRNA) from animal tissues, cells and simple plant tissues using one column
Applications	RT-PCR, qRT-PCR, Northern hybridization, second generation sequencing
Purification method	Mini spin column
Purification technology	Silica technology
Process method	Manual (centrifugation or vacuum)
Sample type	Animal soft tissue, cultured cells, lymphocytes, simple plant tissue
Sample amount	Cells: ≤1 x 107 Animal tissue: 1-20 mgPlant leaves: 50-150 mg Yeast cells: 5 x 106
Yield	2-100μg
Elution volume	≥50μl
Time per run	≤15 minutes（1-24 samples）
Liquid carrying volume per column	800µl
Binding yield of column	100µg

Principle

HiPure RNA technology simplifies total RNA isolation. Samples are first lysed and then homogenized. Ethanol is added to the lysate to provide ideal binding conditions. The lysate is then loaded onto the HiPure silica membrane and RNA binds to the silica membrane, and all contaminants are efficiently washed away. For certain RNA applications that are sensitive to very small amounts of DNA, the residual amounts of DNA remaining can be removed using a convenient on-column DNase treatment. Pure, concentrated RNA is eluted in water.

Advantages

• Efficient removal of DNA – unique genomic DNA removal column without DNase treatment
• High quality – high purity total RNA can be directly used in various sensitive downstream applications
• Fast – several samples can be extracted in 20 minutes by column method
• Safe – no phenol chloroform extraction required
• Sensitive – RNA can be recovered at the level of PG

Kit Contents

Contents	R401102	R401103
Purification Times	50 Preps	250 Preps
HiPure RNA Mini Columns	50	250
2ml Collection Tubes	50	250
RTL Lysis Buffer	50 ml	200 ml
RNA Binding Buffer	15 ml	75 ml
Buffer RW1	50 ml	200 ml
Buffer RW2*	20 ml	2 x 50 ml
RNase Free Water	10 ml	30 ml

Storage and Stability

The Kit can be stored dry at room temperature (15-25°C) and are stable for at least 18 months under these conditions.

The HiPure Total RNA Kit provides fast purification of high-quality RNA from cells, tissues, and yeast using silica-membrane spin columns with a binding capacity of 100μg RNA. There is no need for phenol/chloroform extractions and time-consuming steps such as CsCl gradient ultracentrifugation or isopropanol precipitation. RNA purified using the HiPure Total RNA Purification System can be used for applications such as RT-PCR, Northern blotting, poly A+ RNA (mRNA) purification, nuclease protection, and in vitro translation.

Overview

• Protocol optimized for DNA isolated from a diversity of samples including stool, soil, water, saliva, plant, urine, skin, and more
• Simple and quick workflow: library could be prepared in less than 5 hours
• Component of Norgen’s metagenomics workflow
• A single NGS run can be prepared with up to 384 unique dual-index libraries

Sample type purification kit guide 

The 16S V2-V3 Library Preparation Kit for Illumina consists of the reagents and components required for library preparation of the 16S V2-V3 amplicon libraries to be used for next-generation sequencing on Illumina platforms. All molecular reagents including primers, enzyme mixes, indexes, and buffers are provided. Instructions for PCR clean up with the AMPure XP Magnetic Beads (supplied by customer) are also included for rapid purification of nucleic acid products generated at two steps of the workflow. The library prep workflow could be used for purified DNA inputs from different sources including stool, soil, water, saliva, plant, urine, skin swab, vaginal swab, cheek swab, nasal swab, plasma/serum, tongue swab, gum swab, and others.

The 16S V2-V3 Library Preparation Kit for Illumina has a streamlined procedure that reduces the handling time such that the library prep procedure can be completed in approximately 4 hours (see diagram below). Input DNA is first subjected to targeted PCR to amplify the V2-V3 region of the DNA encoding 16S rRNA. The post-PCR reaction is then cleaned up using AMPure XP beads. Dual index primers are then added using a limited-cycle PCR. The indexed amplicons flanked by 5′ and 3′ barcoded adaptors are then cleaned using AMPure XP beads. The libraries are then ready for quantification, pooling and sequencing.

Details

Supporting Data

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Minimum amount of starting material:	2.5 µL of DNA (5 ng/µL)
Time to complete library preparation:	4 hours

Storage Conditions and Product Stability
Norgen’s 16S V2-V3 Library Prep Kit for Illumina is shipped as one kit box (for the 24 prep kit) or two sub-component kits (for the 96 prep kit). All kits should be stored at -20°C upon arrival.

All kit components should remain stable for at least 1 year when stored at the specified storage conditions.

Step	Component	Cat. 70300 (24 preps)	Cat. 70310, 70320, 70330, 70340 (96 preps)
Amplicon PCR (PCR 1)	MGX Master Mix	330 µL	1,320 µL
	16S V2-V3 Primer Mix	70 µL	280 µL
Index PCR (PCR 2)	Indexing Master Mix	660 µL	2 x 1,320 µL
	N7xx Index Primer	50 µL	50 µL
	S5xx Index Primer	70 µL	70 µL
PCR Clean-Up	Resuspension Buffer	2 x 1,250 µL	2 x 5,000 µL
	Nuclease-free water	1,250 µL	1 x 6,000 µL