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Catalog

17800

Low Copy Plasmid MiniPrep Kit

( 25 preps )
Norgen Biotek

172.06$

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Additional description

Low Copy Plasmid DNA MiniPrep Kit (Cat# 17800 )

For the rapid purification of low copy plasmids

  • High yieldof low copy plasmid DNA
  • Alcohol-free protocol
  • Fast and easy processing

Product Description

Norgen’s Low Copy Plasmid MiniPrep Kit is designed for the rapid preparation of low copy number plasmid DNA from small batch cultures of E. coli. The kit is able to purify 0.5 - 1 µg of low copy number plasmid DNA from 3 mL of bacterial culture. Purification of plasmids up to 14,000 bp in size have been verified.

Purification is based on spin column chromatography using Norgen’s proprietary resin as the separation matrix.Briefly, thebacteriaharbouring the plasmidis pelleted and lysed, and the released plasmid DNA isbound to Norgen's column in the presence of Binding Solution (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 into30 -50µL of the provided ElutionBuffer or water(ELUTE).Pleaseseeprocedure flowchart to the right.

The kit purifies DNA without alcohols, a significant improvement over other techniques to achieve full compatibility with alcohol-sensitive applications. The purified DNA is free of genomic DNA, RNA and cellular proteins and is suitable for many downstream applications.

FEATURES AND BENEFITS

  • Fast and easy processing - Rapid spin-column format allows for the processing of multiple samples in 30 minutes.
  • High yield - 0.5 to 1 µg of low copy number plasmid DNA can be purified from 3 mL of bacterial culture
  • Recovered plasmid DNA is suitable for downstream applications - Purified plasmid DNA is fully compatible with restriction enzyme digestions, and manual orautomated sequencing.
  • Alcohol-free protocol - No alcohols are used during the purificationprocess, thus the DNA is compatible with alcohol sensitive downstream applications.

Specifications

  • Restriction digests
  • Sequencing
  • Transformation

Figure 1.Isolation of High Quality DNA from Low Copy Number Plasmids. The Low Copy Number Plasmid MiniPrep Kit was used to isolate plasmid DNA from 3 mL cultures.Lanes 1-6 are pSYX20 DNA (10-20 copies per cell) while lanes 7-12 are pSC101DNA (2-3 copies per cell).Five microlitres of the total elutions were loaded onto a 1X TAE, 1% agarose gel, with Lanes 1-3 and 7-9 corresponding to 30 µL elutions, and Lanes 4-6 and 10-12 corresponding to 50 µL elutions.In all cases, the DNA is of a high quality, indicated by the presence of sharp bands and no degradation of the isolated DNA.Lane M is Norgen’s MidRanger 1kb DNALadder. Quantification of the DNA is presented in Table 1.

Table 1. High DNA Yield from Small Cultures

The Low Copy Plasmid MiniPrep Kit was used to isolate two different low copy plasmids from 3 mL cultures (as shown in Figure 1). Quantification was performed by loading5µLof the total elutions onto a 1X TAE, 1% agarose gel, followed by densitometry. The average yield of the low copy number plasmids is high in both cases.

APPLICATIONS

Kit Specifications

Column Binding Capacity

25 µg
Average Yield
Up to 1 µg from 3 mL of culture
Size of Plasmids Purified
Up to 14,000 bp
Time to Complete 10 Purifications
30 minutes


LowCopy Plasmid DNA MiniPrep Kit Contents
1.Resuspension Buffer
2.Lysis Solution
3.Binding Solution
4.RNAse A
5.Wash Solution
6.Elution Buffer
7.Micro Spin Columns
8. Collection Tubes
9.Elution Tubes
10.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. The Resuspension Buffer should be stored at 4oC upon addition of RNAse A enzyme.