GenElute™ HP Plasmid Kits

Description: The GenElute™ HP Plasmid Midiprep and Maxiprep Kits offer purification of high-quality plasmid DNA in less than 30 minutes (Figure 1). GenElute HP Plasmid Kits include a filter syringe for lysate clearing, binding columns that can be used with a vacuum or spin purification format, and does not require alcohol precipitation. Fast – high-quality plasmid DNA in 30 minutes Convenient – vacuum or spin column format Cost effective – more value for the money The GenElute HP Plasmid preps deliver significant time-savings and overall higher yields compared to other methods and are Sigma Advanced Technology™ certified. Quick Look: NA0150S, NA0150, NA0160, NA0200S, NA0200, NA0300S, NA0300, NA0310 Procedure: An overnight recombinant E. coli culture is harvested by centrifugation and subjected to a modified alkaline-SDS lysis procedure. The lysate is cleared with a filter syringe and DNA is bound to the silica-based membrane. The remaining contaminates are removed by a wash step. Finally, the bound DNA is eluted in buffer or water. Application: High-quality DNA is ready for immediate use for the most demanding applications, including transfection (Figure 3), restriction digestion (Figure 4), ligation, automated sequencing (Figure 5).

Beneficial Features
Culture Volume	1-5 ml (Mini), 50 ml (Midi) and 150 ml (Maxi) bacterial culture
Yield	Up to 25 µg (Mini), 350 µg (Midi) and 1.2 mg (Maxi) yield of high-copy   plasmid DNA (Figure 2)
Speed	From harvested bacterial culture to pure plasmid DNA in 30 minutes or less
Flexible	Offers the flexibility of a vacuum or spin format; does not rely on gravity flow   or ultracentrifugation
Economic	Lower cost per prep plus higher yields per prep equals double savings

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Order Information
GenElute HP Plasmid DNA Kits
Product No.	Product Name	Purifications per kit	Technical Manual .pdf	Short Protocol .pdf
NA0150S	GenElute HP Plasmid Miniprep Kit	10	X	-
NA0150	GenElute HP Plasmid Miniprep Kit	70	X	-
NA0160	GenElute HP Plasmid Miniprep Kit	350	X	-
NA0200S	GenElute HP Plasmid Midiprep Kit	4	X	X
NA0200	GenElute HP Plasmid Midiprep Kit	25	X	X
NA0300S	GenElute HP Plasmid Maxiprep Kit	4	X	X
NA0300	GenElute HP Plasmid Maxiprep Kit	10	X	X
NA0310	GenElute HP Plasmid Maxiprep Kit	25	X	X

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Sample Data

Preparation Time and Plasmid DNA Yield for different Maxiprep Kits

Figure 1. Comparison of time required per isolation using different Maxiprep plasmid isolation systems. Plasmid DNA was isolated following each manufacturers recommended protocol. Each system was tested multiple times and the average preparation time is shown here.   Figure 2. Comparison of plasmid DNA yield using different Maxiprep plasimd isolation systems. Plasmid DNA was isolated following each manufacturers recommended protocol. All samples were done in duplicate.

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Figure 3. Comparison of transfection efficiencies of plasmids prepared with different plasmid isolation methods. Data shows an average of 3 replicates for each named isolation method. All transfections were in CHO-K1 cells. The OD measurements were taken at 420 nm recording the units of ß-gall/plate. The cells were grown to 60-70% confluency and transfected using 3 µg of plasmid pSPORT-CMV/15 µg of Escort IV (Product No. L3287). b-Gal activity was determined 54 hours post-transfection.   Figure 4. Restriction digestion of plasmid DNA isolated using the GenElute HP Plasmid Maxiprep Kits and a comparable kit from Supplier Q. The plasmid DNA (800 ng) was digested with 10 units of Hind III at 37 °C for 4 hours. Each plasmid sample represents 100 ng of undigested DNA (left) and digested DNA (right) loaded in each pair of lanes. Samples were loaded on a 1% TBE agarose gel. The marker (M) used was 1 kb DNA Ladder (Product No. D0428).

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High-quality DNA for Automated Sequencing

Figure 5. Electropherogram revealing >600 bases of sequence from pCMV-SPORT-b-gal purified from GenElute HP Plasmid Midiprep Kit. Cycle Sequencing was performed using 500 ng template, a T7 sequencing primer and ABI BigDye™ terminator chemistry. Sequencing reactions were resolved on an ABI Prism XL instrument with a 48 cm gel cassette containing 4.5% AutoPAGE Plus acrylamide at 2.4 kV, 48°C for 7 hours.

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