Low Copy Plasmid DNA MiniPrep

Low Copy Plasmid DNA MiniPrep

Catalog# : NB-17800
Price/Unit : $110.00
Unit : 25 preps
For the rapid purification of low copy plasmids

* High yield of low copy plasmid DNA
* Alcohol-free protocol
* Fast and easy processing

Bio-Synthesis'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 Bio-Synthesis's proprietary resin as the separation matrix. Briefly, the bacteria harbouring the plasmid is pelleted and lysed, and the released plasmid DNA is bound to Bio-Synthesis's column in the presence of Binding Solution (BIND). Under these conditions only the DNA will bind to Bio-Synthesis's resin while most of the contaminating RNA and cellular proteinaceous components are removed in the flowthrough. The bound DNA is then washed to remove any remaining impurities (WASH). Lastly, the purified DNA is eluted into 30 - 50 µL of the provided Elution Buffer or water (ELUTE). Please see procedure 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 or automated sequencing.
* Alcohol-free protocol - No alcohols are used during the purification process, thus the DNA is compatible with alcohol sensitive downstream applications.


APPLICATIONS

* 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 Bio-Synthesis's MidRanger 1kb DNA Ladder. Quantification of the DNA is presented in Table 1.

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

Long-Term Storage : 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.
Product Usage
For laboratory research use only, not for diagnostic use! Not for use in humans.