GeneAb™ Kappa

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

• Real time qPCR kit
• For screening Anatoxin gene cluster
• Use in combination with Attogene Algae DNA isolation kit

Description

Not all cyanobacterial strains produce toxins. However, the toxin-producing strains cannot be distinguished from the nontoxin-producing strains by traditional light microscopy, commonly
used to monitor water bodies.  An alternative for the differentiation of potentially toxic strains from nontoxic strains is to use molecular methods to detect the presence of toxin biosynthetic genes. Such methods are already available and could be used for the detection and identification of potential microcystin and nodularin producers present in environmental samples (Attogene catalog number NA2024).

Screening for the toxin itself, can be very costly.  In turn, real time PCR for the detection of the anaC gene in cyanobacterial strains and environmental samples can be a key indicator for the prescense of cyanobacteria capable of expressing the Anatoxin toxin.  Attogen has thus, designed primer pairs and probes targeting a the conserved anaC gene region in order to enable the amplification and detection of several producer genera using real time PCR.  Screening for the toxin genes can save significant costs and act as a triage for samples needing to be analyzed for the toxin itself.

Real time qPCR kit
For screening Anatoxin gene cluster
Use in combination with Attogene Algae DNA isolation kit

The NGS Cell Free DNA Library Prep Kit (illumina and MGI Platforms) was developed for the construction of high quality cell-free DNA (cfDNA) libraries using 1 ng to 50 ng of cell-free DNA as input. The kit has a simple work flow and a fast procedure. Multiplexing of the cell free DNA library is possible based on the index type.

NGS Cell Free DNA Library Prep Kit Workflow

The main source of cell-free DNA (cfDNA) is derived from apoptotic hematopoietic cells in blood and found in the plasma. The length of the cfDNA is about 150-200 bp in length. Circulating tumor DNA (ctDNA) derived from malignant tumors is a part of cfDNA. Both cfDNA and ctDNA can be used as a noninvasive biomarker since it offers a better approach in comparison to invasive tissue biopsies.

NGS has been used for cfDNA and ctDNA sequencing in the field of liquid biopsy as it provides a whole genome level of molecular profiling. One of the hurdles for cfDNA sequencing is the difficulty of library preparation from the limited amount of cfDNA obtained from plasma. Our cell-free NGS kit makes it easy to get enough libraries from limited input in just 1.5 hours.

Three index types are available for the NGS Cell Free DNA Library Prep Kit of the illumina platform:

Non-index (Cat.# 30029): Libraries do not have index.

Index(Cat.# 30031): Each of our index primers contains a unique 6-base index sequence that can be used for sample identification. Total 48  library multiplexing is possible. Index information can be downloaded here.

Unique dual index (Cat.# 30033): Cell-free DNA library multiplexing up to 96 samples is possible with the unique dual indexes. We have developed a Four-Base Difference Index System. The system have at least 4 bases different from each other in the 8 bases index length. The primers effectively minimize sequencing errors such as mis-assignment, index hopping, index contamination etc. Index information can be downloaded here.

Indexes are available for the MGI platform kits (Cat.# 34031).

Kit advantages:

• Fast and simple protocol
  • Hands-on time is around 10 minutes
  • The total protocol time is only 1.5 hours
• Easy procedure based on:
  • Ready-to-use master mix for easy setup of reactions
  • Less reaction components to simplify bench work
• Less magnetic beads used for cleanup procedures: This can reduce more than 50% of the beads consumption
• Guaranteed high library conversion efficiency
• Low input cell-free DNA: From 1 ng to 50 ng

Comparison of library conversion efficiency with different samples under the same condition.

Comparison of library yield with different samples under the same condition.

The NGS Cell Free DNA Library Prep Kit (illumina and MGI Platforms) was developed for the construction of high quality cell-free DNA (cfDNA) libraries using 1 ng to 50 ng of cell-free DNA as input. The kit has a simple work flow and a fast procedure. Multiplexing of the cell free DNA library is possible based on the index type.