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Catalog

13100

DNA Gel Extraction Kit

( 50 preps )
Norgen Biotek

199.25$

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Additional description

Direct Fungi DNA Isolation Kit (Cat# 25600 )

For the rapid and convenient isolation of fungal DNA from plant surfaces

  • Rapid isolation of fungal DNA from plantsurfaces
  • No liquid nitrogen or physical disruption methods required
  • Isolate high quality DNA for a wide range of downstream applications

Product Description

Norgen’s Direct Fungi DNA Isolation Kit provides a rapid method for the isolation and purification of total DNA from the fungi found on the surface of symptomatic plant samples. The major benefit of this kit is that it does not require the use of physical homogenization steps to isolate the fungal DNA, which are often time consuming and tedious procedures that are not suitable methods for use in the analysis of large numbers of samples. Furthermore, in the area of pathogen diagnosis simple and convenient sample preparation steps are a critical factor in supporting high throughput analysis. Therefore, Norgen’s Direct Fungi DNA Isolation Kit adopts a simple and rapid sample preparation procedure that does not rely on physical homogenization steps to isolate sufficient amounts of high quality fungal DNA that can be used in sensitive downstream detection methods.

Purification is based on spin column chromatography using Norgen's proprietary resin as the separation matrix. Briefly, the surface fungus is extracted from the plant tissue of interest by shaking with a small volume of Collection Solution. The collected fungi are then lysed, and the released fungal DNA is bound to Norgen's column(BIND).Under these conditions only the DNA will bindto Norgen's resin whilemost ofthe contaminating RNAand cellular proteinaceous componentsare removed in the flowthrough. The bound DNAis then washed to remove anyremaining impurities(WASH).Lastly, the purified DNA is eluted into 50 - 100µL of the provided ElutionBuffer or water(ELUTE).Pleaseseeprocedure flowchart to the right.

FEATURES AND BENEFITS

  • No liquid nitrogen or physical disruption methods required- Rapidly isolate fungal DNA from the surface of symptomatic plant samples without any physical disruption methods.
  • Rapid and simple processing- Rapid spin-column format and no physical disruption allows for the processing of multiple samples in 60 minutes.
  • High quality fungal DNA- The purified DNA is of the highest quality and can be used in a number of sensitive downstream applications, including PCR.
  • Purified DNA can be use in a number of downstream applications- Purified fungal DNA can be used in a number of downstream applications including real time PCR, sequencing, Southern blotting and SNP analysis.

Specifications

  • PCR
  • qPCR
  • Sequencing
  • Southern Blot Analysis
  • SNP Analysis

Figure 1. PCR Amplification of Fungal Pathogens from Grapes for Sequencing Analysis. Total DNA was isolated from the surface of two symptomatic grapes usingNorgen's Direct Fungi DNA Isolation Kit. Next, 2 µL of the 50 µLelutionswere subsequently used as the templatein end-point PCR reactions using universal primers for the fungal ITS region. For analysis 10 µL of the PCR products were loaded on the 1% agarose gel above. The bands were extracted by Norgen’s DNA Gel Extraction Kit (Cat. 13100 ) for sequencing analysis. Through sequencing it was determined that DNA was successfully isolated and amplified from 7 different fungal species including A. niger (Lane 1), C. cladosporiodes (Lane 2), B. cinerea (Lane 3), M. racemosus (Lane 4), A. tenuissima (Lane 5), Penecillium sp. (Lane 5), F. oxysporum (Lane 6) and R. oryzae (Lane 7). Lane C is the non-template control and Lane M is Norgen’s PCRSizer DNA Ladder.

Figure 2. Specific Detection of Botrytis cinerea Infection using Real-time PCR. Fungal DNA was isolated from 5 different varieties of symptomatic grape berries randomly sampled in a vineyard usingNorgen's Direct Fungi DNA Isolation Kit. The isolatedDNA was then used in a real-time PCR reaction for the specific detection of Botrytis cinerea infection. Briefly, 2 µL of DNA from each 50 µL elution was mixed in 20 µL of total PCR reaction buffer and real-time PCR was performed. From observing the graph above, thereal-time PCR was able to identify Botrytis infection from the Cabernet Sauvignon and the Viognier grapes, indicate that the quality of DNA isolated from Direct Fungi DNA Isolation Kit is suitable for any downstream application.

APPLICATIONS

Kit Specifications
Column Binding Capacity
15 µg
Maximum Column Loading Volume
650 µL
Maximum Amount of Starting Material:
Symptomatic Grapes
Plant Tissue
1 – 2 berries (<1 gram total weight)
<1 gram total weight
Time to Complete 10 Purifications
60 minutes


Direct Fungi DNA Isolation Kit Contents
1. Resuspension Buffer
2. Lysis Solution
3. Wash Solution I
4. Wash Solution II
5. Elution Buffer
6. Mini Spin Columns
7. Collection Tubes
8. Elution Tubes
9. Product Insert


Storage Conditions and Product Stability
All solutions should be kept tightly sealed and stored at room temperature. These reagents should remain stable for at least 1 year in their unopened containers.