Eppendorf provides a complete system for long-range PCR1 analysis of gDNA. The Perfect gDNA Blood Mini Kit offers an easy and effective method for isolating high molecular weight gDNA. The resulting gDNA can be used in combination with the Eppendorf Mastercycler gradient thermocycler and Eppendorf TripleMaster PCR System to amplify long templates.
In the past, scientists needing pure, high molecular weight gDNA would have to purchase special products or use lengthy, highly involved methods for DNA isolation. The Perfect gDNA Blood Mini Kit provides a method of extraction resulting in DNA ranging in size from 20 kb to 90 kb. PCR fragments up to 24 kb in size have been amplified using gDNA purified by the Eppendorf method. The following set of experiments was designed to demonstrate how Perfect gDNA Blood Mini Kit and TripleMaster team up to enable easy amplification of long templates.
Materials and Methods
Pulse Field Gel Electrophoresis:
A number of samples isolated using the Perfect gDNA Blood Mini Kit (Eppendorf) and QIAamp DNA Blood Mini Kit (Qiagen, Hilden, Germany) were analyzed using pulse field gel electrophoresis (see Fig. 1). Samples were loaded on a 1% agarose gel in 0.5X TBE Buffer using the BIO-RAD Pulse Field Electrophoresis apparatus (BIO-RAD, Hercules, CA, USA). A Low Range PFG Marker (NEB, Beverly, MA, USA) and Lambda Hind III Marker (Worthington, Lakewood, NJ, USA) were run with the samples for size determin