Introduction

This product is suitable for rapid extraction of total DNA from tissue, cells, blood, saliva, swabs, blood spots, semen and other clinical samples. DNA can be used directly for PCR, quantitative PCR, Southern Blot, test of virus DNA and so on.

Details

Specifications

Principle

This product is based on silica Column purification. The sample is lysed and digested with lysate and protease, DNA is released into the lysate. Transfer to an adsorption column. Nucleic acid is adsorbed on the membrane, while protein is not adsorbed and is removed with filtration. After washing proteins and other impurities, Nucleic acid was finally eluted with low-salt buffer (10mm Tris, pH9.0, 0.5mm EDTA).

Advantages

• High quality DNA – meet a variety of downstream applications, including PCR, qPCR, enzyme digestion, hybridization, etc.
• Fast – without separation of leukocytes, organic extraction or ethanol precipitation
• Simple – all nucleic acids can be obtained by direct digestion
• Wide applicability- handle a variety of liquid samples

Kit Contents

Storage and Stability

Proteinase K should be stored at 2-8°C upon arrival. However, short-term storage (up to 12 weeks) at room temperature (15-25°C) does not affect their performance. The remaining kit components can be stored at room temperature (15-25°C) and are stable for at least 18 months under these conditions. The entire kit can be stored at 2-8°C, but in this case buffers should be redissolved before use. Make sure that all buffers are at room temperature when used.

Experiment Data

This product is suitable for rapid extraction of total DNA from tissue, cells, blood, saliva, swabs, blood spots, semen and other clinical samples. DNA can be used directly for PCR, quantitative PCR, Southern Blot, test of virus DNA and so on.

Description 

Champion™ E. coli Transformation Kit provides an easy method for rapid preparation of chemically competent cells with high transformation efficiency from fresh culture, overnight culture, or even directly from bacterial colonies on the plate. The competent cell preparation method eliminates the requirement of time-wasting wash step. In addition, preparation of competent cells from overnight culture or directly from bacterial colonies provides flexibility to cloning experiment. The resultant competent cells can be immediately used or stored at -70°C for one year. This kit includes a specialized SMO-Broth™ medium and a unique Champion™ CC Buffer for culturing and preparing competent cells efficiently. Following the simple and quick competent cell preparation protocol from fresh culture, the transformation efficiency is typically ranged from 10⁸–10⁹ cfu/μg transformants/μg of pUC19 plasmid DNA, but varies depending on the E. coli strains. The resultant competent cells can be further transformed using time-saving transformation protocol, eliminating the requirement of heat-shock and recovery steps. 

Features

• Flexible– fresh culture, overnight culture, 4°C stored liquid culture or even colonies on agar plate can be used for transformation. 
• Fast and Easy– only few steps for preparation; suitable for time-saving transformation 
• High efficiency– up to 109 cfu/μg 
• Personalization– suitable for most E. coli strains

Kit Contents

Storage

4°C for 12 months

Champion™ E. coli Transformation Kit provides an easy method for rapid preparation of chemically competent cells with high transformation efficiency from fresh culture, overnight culture, or even directly from bacterial colonies on the plate. The competent cell preparation method eliminates the requirement of time-wasting wash step. In addition, preparation of competent cells from overnight culture or directly from bacterial colonies provides flexibility to cloning experiment. The resultant competent cells can be immediately used or stored at -70°C for one year. This kit includes a specialized SMO-Broth™ medium and a unique Champion™ CC Buffer for culturing and preparing competent cells efficiently. Following the simple and quick competent cell preparation protocol from fresh culture, the transformation efficiency is typically ranged from 108–109 cfu/μg transformants/μg of pUC19 plasmid DNA, but varies depending on the E. coli strains. The resultant competent cells can be further transformed using time-saving transformation protocol, eliminating the requirement of heat-shock and recovery steps. 

K-RINTDF

SKU: 700004335

100 assays per kit

The Rapid Integrated Total Dietary Fiber Assay Kit method is validated under collaborative study (AACC Method 32-60.01, AOAC Method 2022.01, AOAC Method 2017.16, ICC Standard No. 185) and is recognized as a Type I Method by CODEX Alimentarius. The K-RINTDF method is the recommended one for the measurement of total dietary fiber in all foods that may or may not contain resistant starch. This method is updated to be more consistent with in vivo conditions in the human small intestine, i.e. a 4 h incubation time. Under these conditions more accurate measurement of resistant starch is obtained, including phosphate cross-liked starch (RS₄). Use of higher enzyme concentrations ensures that resistant maltodextrins produced from non-resistant starch under the incubation conditions of the Integrated Total Dietary Fiber procedure (AOAC Methods 2009.01 and 2011.25) are no longer produced.

In this improved, rapid method, the incubation time with PAA + AMG is reduced to 4 h and the levels of both PAA and AMG are increased to ensure that resistant starch levels obtained with a set of control samples are consistent with ileostomy data. Under these conditions, the DF values obtained for most samples are the same as those obtained with AOAC Methods 2009.01 and 2011.25.

The dietary fiber fractions that are measured with this method are:

1. High Molecular Weight Dietary Fiber (HMWDF) including Insoluble Dietary Fiber (IDF) and High Molecular Weight Soluble Dietary Fiber (SDFP; soluble dietary fiber which is precipitated in the presence of 78% aqueous ethanol), and

2. Low Molecular Weight Soluble Dietary Fiber (SDFS; water soluble dietary fiber that is soluble in the presence of 78% aqueous ethanol).

Alternatively, IDF, SDFP and SDFS can be measured separately.

The enzymes used in this method are high purity and effectively devoid of contaminating enzymes active on other dietary fiber components such as β-glucan, pectin and arabinoxylan. They are supplied as freeze-dried powders; allowing the use of glycerol as an internal standard in the method.

See our full range of dietary fiber assay kits.

View Dietary Fiber Measurement Guide – Which Method for which sample?

* See McCleary, B. V., Sloane, N & Draga, A. (2015). Determination of total dietary fibre and available carbohydrates: a rapid integrated procedure that simulates in vivo digestion. Starch/Starke, 66, 1-24.

Validation of Methods

Advantages

• More rapid measurement – incubation time with PAA + AMG reduced to 4 h in comparison with AOAC 2009.01 (increased levels of enzyme employed) 
• DF values for most samples are very similar to those obtained with AOAC Method 2009.01 
• Rapid Integrated Total Dietary Fiber method removes all of the limitations that have been identified with AOAC Method 2009.01* 
• All reagents stable for > 2 years after preparation 
• The method is consistent with the CODEX Alimentarius definition of dietary fiber 
• Mega-Calc™ software tool is available from our website for hassle-free raw data processing 
• Very competitive price (cost per test)

The Rapid Integrated Total Dietary Fiber Assay Kit method is validated under collaborative study (AACC Method 32-60.01, AOAC Method 2022.01, AOAC Method 2017.16, ICC Standard No. 185) and is recognized as a Type I Method by CODEX Alimentarius. The K-RINTDF method is the recommended one for the measurement of total dietary fiber in all foods that may or may not contain resistant starch. This method is updated to be more consistent with in vivo conditions in the human small intestine, i.e. a 4 h incubation time. Under these conditions more accurate measurement of resistant starch is obtained, including phosphate cross-liked starch (RS4). Use of higher enzyme concentrations ensures that resistant maltodextrins produced from non-resistant starch under the incubation conditions of the Integrated Total Dietary Fiber procedure (AOAC Methods 2009.01 and 2011.25) are no longer produced.