Catalog

11900

HighRanger 1kb DNA Ladder

( 100 loads )
Norgen Biotek

172.06$

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Additional description

Phage DNA Isolation Kit (Cat# 46700 )

For the rapid purification of total DNA from bacteriophages

  • Isolate high quality DNA from a variety of phage strains
  • High yields of total DNA
  • Fast and easy processing using a rapid spin-column format
  • No phenol or chloroform extractions or cesium chloride banding required

Product Description

Norgen’s Phage DNA Isolation Kit provides a rapid method for the isolation and purification of total DNA from bacteriophages propagated in bacteria grown in liquid cultures. The DNA is isolated without the use of phenol, chloroform or cesium chloride. The spin-column based procedure is rapid, and can be completed in less than 45 minutes. The kit is highly efficient for processing small volumes of phage supernatant (500 µL – 1 mL).

The starting material for this procedure is clarified phage supernatant that has been purified from bacteria grown either in liquid cultures. Briefly, the phage particles are lysed through a heat and chemical lysis process with the provided Lysis Solution. The released phage DNA is then bound to Norgen's spin column in the presence of BindingSolution (BIND). Under these conditions only the DNA will bind tothe columnwhile most of the RNA and proteins are removed in the flowthrough. The bound DNA is then washed to remove any remaining impurities (WASH). Lastly, the purified total phage DNA is eluted into 50 - 100µLof the provided Elution Buffer or water (ELUTE). Please see the flowchart to the right. The purified total phage DNA is of the highest integrity, and can be used in a number of downstream applications.

FEATURES AND BENEFITS

  • Fast and easy processing - Rapid spin-column format allows for the processing of multiple samples in45 minutes
  • Simple procedure - Isolate total phage DNAusing a rapid spin-column format; nophenol extractions or cesium chloride banding required
  • Isolate total phage DNAfrom a variety of phage strains
  • Recovered genomic DNA is suitable for downstream applications - Purified genomic DNA is fully compatible with restriction enzyme digestions, Southern Blot and PCR analysis.
  • Highyield of totalDNA - Isolate10-30 µg DNA from 106-1010 pfu/ mL of enriched phages

Specifications

Purified total phage DNA is highly suited to a wide range of applications including

  • Restriction enzyme digestions
  • PCR
  • Southern blot analysis

Figure 1. Effective Host Genomic DNA Removal without Reducing Phage DNA Yield. Total DNA was isolated from four enriched phage cultures using Norgen’s Phage DNA Isolation Kit. A DNase I pre-treatment was performed prior to adding the provided Lysis Solution. Briefly, 20 units of DNase I was added to 1 mL of enriched phage culture and the mixture was incubated at room temperature for 20 minutes. After the DNAase I treatment the procedure was followed. As a control, DNA was isolated from aliquots of the same 4 cultures using Norgen’s Phage DNA Isolation Kit without performing the DNase I treatment. For DNA analysis 10 μL of each 50 μL elution was loaded onto a 1X TAE agarose gel. As it can be seen, the phage DNA was safely protected from the DNase I treatment by its coat protein, while the host genomic DNA was efficiently degraded by the DNase I. Thus the DNase I pre-treatment resulted in less host gDNA contamination in the final phage elution without influencing the total phage DNA yield. Lane M is Norgen’s Highranger 1 kb DNA Ladder (Cat. 11900)

Figure 2. Optional Proteinase K Treatment Improves DNA Yield for Certain Phage Strains. Total DNA was isolated with and without the optional Proteinase K treatment using Norgen’s Phage DNA isolation kit. Briefly, 4 µL of Proteinase K (20 mg/mL) was added to 1 mL of enriched phage culture and incubated at 55°C for 15 minutes with a the phage Lysis Buffer. After the Proteinase K treatment the procedure was followed. As a control, DNA was isolated from aliquots of the same 8 cultures using Norgen’s Phage DNA Isolation Kit without performing the Proteinase K treatment. For DNA analysis 10 μL of each 50 μL elution was loaded onto a 1X TAE agarose gel and the yield of DNA was compared from the eight different phage types (lane 1 to 8). As it can be seen, the optional treatment of Proteinase K improved the phage DNA yield in Lanes 2, 5 and 6 dramatically. Lane M is Norgen’s Highranger 1 kb DNA Ladder (Cat. 11900)

APPLICATIONS

Kit Specifications
Column Binding Capacity
50 µg
Maximum Column Loading Volume
650 µL
Size of DNA Purified
All sizes
Maximum Amount of Starting Material
1 × 1010 pfu/mL enriched phages
Average Yield*
3-15 µg DNA from 106-1010pfu/ mL of enriched phages
Time to Complete 10 Purifications
45 minutes
*Average yields will vary depending upon a number conditions used and developmental stage.

Phage DNA Isolation Kit Contents
Lysis Solution
Binding Solution
Wash Solution
Elution Buffer
Spin Columns inserted into Collection Tubes
Elution tubes (1.7 mL)
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.