GenElute™ HP 96 Well Plasmid Miniprep Kit

Plasmid DNA Purification High-throughput purification of plasmid DNA in less than 50 minutes

The GenElute HP 96-Well Plasmid Miniprep Kit is designed for high-throughput purification of plasmid DNA from bacterial cultures. The kit protocol can be performed manually with the use of a vacuum manifold and is also compatible with automated liquid handling workstations. The kit is capable of generating a yield of up to 10 µg of high-copy plasmid DNA with a protocol that can be completed in under 50 minutes. Designed to complement Sigma’s MISSION® shRNA product line, the GenElute HP 96-Well Plasmid Miniprep Kit is optimized for the most demanding downstream transfection protocols (Fig. 1). The result is purified MISSION® shRNA plasmid DNA that can generate best-in-class viral titers when compared to plasmids purified with competitive products (Fig. 2). In addition, the kit generates purified plasmid DNA suitable for a wide variety of molecular biology applications, including restriction digestions (Fig. 3) and sequencing (Fig. 4). Features and Benefits From harvested bacterial culture to pure plasmid DNA in under 50 minutes Up to 10 µg yield of high-copy plasmid DNA per well Optimized for best-in-class production of viral titers from purified MISSION® shRNA plasmid vectors Generates purified plasmid DNA for a wide variety of downstream applications, including transfections, sequencing, and restriction digests Offers the flexibility of both manual and automated formats No phenol/chloroform extraction or alcohol precipitation required Kits are stable at room temperature for convenient storage

Purified plasmid DNA is suitable for transfection experiments Figure 1. Cells expressing TurboGFP™ 40 hours post-transfection. HEK293T cells were seeded at 20,000 viable cells/well in a 96-well plate. 24 hours post-seeding, cells were transfected with MISSION® TurboGFP Control Vector (SHC003) purified with the GenElute HP 96 Well Plasmid Miniprep kit. The cell culture medium was replaced at 16 hours and 40 hours post-transfection and then imaged using a fluorescent microscope.

Robust viral titers from purified MISSION® shRNA plasmid DNA clones Figure 2. Titer measurements of lentiviral particles from cells transfected with a purified MISSION® shRNA plasmid DNA clone. MISSION® shRNA plasmid DNA was isolated according to the protocol of each supplier. The purified plasmid DNA was then used to transfect HEK293T cells. The titer measurements of the resulting lentiviral particles were then determined using a p24-based ELISA. The final data represents the average of 12 experimental replicates for each manufacturer.

Purified plasmid DNA is suitable for restriction enzyme digestion Figure 3. Restriction digestion of plasmid DNA isolated with the GenElute™ HP 96-Well Plasmid Miniprep kit. MISSION® shRNA Plasmid DNA was purified in a 96-well format. 40 µl PvuII digests were set up using 4 µl from each recovered plasmid. The DNA was digested for 1 hour at 37 °C and then loaded onto the Caliper LabChip® instrument. The LabChip instrument analyzed the DNA and provided a visual image showing the correct digestion pattern for all samples.

High-quality DNA for automated sequencing

Figure 4. Electropherogram showing purified plasmid DNA sequence. pLKO.1-puro plasmid DNA was purified using the GenElute HP 96-Well Plasmid Miniprep kit. The product was then sequenced using BigDye® terminator v3.1 chemistry. Sequencing reactions were analyzed on an ABI 3730xl.

Ordering Information

Product No.	Product Description	Size	Quantity
NA9604	GenElute HP 96 Well Plasmid Miniprep Kit	4 × 96	1 Kit

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