α-Amylase Reagent (Ceralpha)

Overview

• Rapid isolation of both small and large species of DNA from urine
• Convenient spin column format
• Effective removal of PCR inhibitors
• Purified DNA is highly suited to sensitive downstream applications
• Allows for the purification of viral DNA from urine

Both high molecular weight DNA (greater than 1 kb in size; mostly cell associated) and the smaller DNA (150 – 250 bp; derived from the circulation) is effectively isolated and purified using a rapid and convenient spin column protocol. This kit can be used to isolate DNA from a broad range of viruses in urine as well. Salts, metabolic wastes, proteins and other contaminants are removed to yield inhibitor-free DNA for use in sensitive applications. The DNA is of excellent quality for various downstream applications such as PCR, qPCR and DNA fingerprinting, methylation studies and more.

This kit is fully compatible with Norgen’s Urine Collection and Preservation Tubes.

Urine DNA Isolation Kit

This kit provides a fast, reliable and simple procedure for isolating DNA from urine volumes ranging from 50 μL to 1.75 mL of urine. Multiple samples can be processed in 30 minutes.

Urine DNA Isolation Kit (Slurry Format)

This kit provides a fast, reliable and simple procedure for isolating DNA from urine volumes ranging from 3 mL to 25 mL. Multiple samples can be processed in 30 minutes. Multiple samples can be processed in 45 minutes.

Urine DNA Isolation Maxi Kit (Slurry Format)

This kit provides a fast, reliable and simple procedure for isolating DNA from urine volumes ranging from 25 mL of urine up to 80 mL. Multiple samples can be processed in 45 minutes.

Background

DNA found in urine can be divided into 2 basic categories. The larger species, genomic-DNA (gDNA), is generally greater than 1 kb in size, and appears to be derived mainly from exfoliated cells. The second species is smaller, generally between 150 and 250 bp (apoptotic-DNA), and derives, at least in part, from the circulation. The second species is also considered as an RNA/DNA hybrid as reported by Halicka et al. (2000). Both types of DNA can be isolated reliably using this kit.

Details

Supporting Data

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Kit Specifications
Volume of Urine Processed	1.75 mL
Average Yield*	Up to 50 ng
Size of DNA Purified	Large (> 1 kb) and small (150-250 bp)
Time to Complete 10 Purifications	30 minutes hands-on time (plus a 1 hour incubation)

* Yield will vary depending on the type of sample processed


Storage Conditions and Product Stability
All buffers should be kept tightly sealed and stored at room temperature. This kit is stable for 2 years after the date of shipment. The kit contains a ready-to-use Proteinase K, which is dissolved in a specially prepared storage buffer. The buffered Proteinase K is stable for up to 2 years after the date of shipment when stored at room temperature.

Component	Cat. 18100 (50 preps)	Cat. 48800 (50 preps)	Cat. 50100 (50 preps)
Binding Solution K	15 mL	–	–
Slurry B1	–	18 mL	18 mL
Proteinase K in Storage Buffer	2 mL	–	–
Pronase K in Storage Buffer	2 mL	–	–
Soluton WN	9 mL	–	–
Binding Buffer A	–	–	50 mL
Lysis Buffer A	–	30 mL	30 mL
Wash Solution A	–	38 mL	38 mL
Wash Solution B	30 mL	–	–
Wash Solution D	9 mL	–	–
Binding Solution K	15 mL	–	–
Elution Buffer B	15 mL	15 mL	15 mL
Micro Spin Columns	50	–	–
Mini Filter Spin Columns	–	50	50
Collection Tubes	50	50	50
Elution Tubes (1.7 mL)	100	100	100
Product Insert	1	1	1

Overview

• All-in-one solution for inclusion body protein isolation and purification
• Fast and convenient spin column protocol
• Complete kit with Cell Lysis Reagent, Inclusion Body Solubilization Reagent, buffers and spin columns to purify proteins
• Purification is based on spin column chromatography that uses Norgen’s resin separation matrix

These kits provide everything required to isolate and purify inclusion body proteins from induced bacterial cultures. First a proprietary Cell Lysis Reagent is used to selectively lyse the cells and release inclusion bodies in their solid form. Next, inclusion bodies are dissolved and their contents released using the provided IB Solubilization Reagent. Inclusion body proteins are then further purified using spin columns for rapid and convenient buffer exchange and desalting. This kit provides a convenient way to screen recombinants prior to scaling up.

ProteoSpin™ Inclusion Body Protein Isolation Micro Kit

The process is efficient and streamlined and can process up to 12 samples in only 60 minutes. Each spin column is able to recover up to 50 µg of acidic or basic proteins. Purified recombinant proteins are then ready for SDS-PAGE, 2D gels, Western blots, Mass Spectrometry analysis, and other applications.

ProteoSpin™ Inclusion Body Protein Isolation Maxi Kit

The procedure is efficient and streamlined and can process up to 4 samples in approximately 2 hours. Each spin column is able to recover up to 12 mg of acidic or basic proteins from 100 mL of induced bacterial culture. Purified recombinant proteins are then ready for SDS-PAGE, 2D gels, Western blots, Mass Spectrometry analysis, and other applications.

About Inclusion Bodies

Bacteria are widely used for the expression of different proteins. However, 70-80% of the proteins expressed in bacteria by recombinant techniques are typically contained in insoluble inclusion bodies (i.e., protein aggregates). The protein of interest found in these subcellular structures is often inactive, due to incorrect folding. The production rate of recombinant proteins stored in inclusion bodies is invariably higher than those synthesized as soluble proteins. The reason behind this is thought to be the resistance of insoluble proteins to proteolysis by cellular enzymes. In addition, separation of insoluble recombinant proteins in inclusion bodies is considerably easier than that of soluble proteins. These factors have been the major influences favoring scale-up of high-value proteins using bacterial fermentation for example. Procedures for the purification of the expressed proteins from inclusion bodies are often labour-intensive, time-consuming and not cost-effective. This kit provides the essential reagents for cell disruption, inclusion body solubilization and purification using spin column chromatography – all optimized to work together thereby simplifying the process and saving a tremendous amount of time and cost.

Details

Supporting Data

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Kit Specifications
Maximum Culture Volume	100 mL
Yield from 100 mL of Culture	Up to 12 mg
Minimum Elution Volume	4 mL
Time to Process 1 Sample	1 hour

Storage Conditions
The Cell Lysis Reagent and IB Solubilization Reagent should be stored at 4°C upon receipt of this kit. This kit is stable for 2 years after the date of shipment. Once opened, the solutions should be stored at 4°C when not in use except for Binding Buffer C and Binding Buffer N. Some precipitation will occur with 4°C storage. This precipitation should be dissolved with slight heating to room temperature before using.

Component	Cat. 10300 (Micro – 25 preps)	Cat. 17700 (Maxi – 4 preps)
Wash Solution C	30 mL	130 mL
Wash Solution N	30 mL	130 mL
Binding BUffer A	4 mL	20 mL
Binding Buffer N	4 mL	20 mL
Elution Buffer C	8 mL	2 x 30 mL
Protein Neutralizer	4 mL	4 mL
Cell Lysis Reagent	15 mL	110 mL
IB Solubilization Reagent	2 mL	50 mL
Syringes, 1cc, slip tip	25	–
Needles (Bev, 20G x 1 inch)	25	–
Syringes, 10 mL, Luer-Lok™ Tip	–	4
Needles (18G x 1.5 inch)	–	4
Micro Spin Columns	25	–
Maxi Spin Columns (filled with SiC) inserted into 50 mL collection tubes	–	4
Collection Tubes	25	–
Elution Tubes (1.7 mL)	25	–
Elution Tubes (50 mL)	–	4
Product Insert	1	1