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Generation Capture Plate Kit

For quick high-throughput purification of DNA for standard PCR
  • Fast and easy protocol
  • Well-suited for standard PCR screening applications
  • Purified DNA is ready to use in standard PCR

The Generation Capture Plate Kit provides the components and procedures necessary for purifying genomic DNA, in a 96-well format, from whole blood, bone marrow, buffy coat, body fluids, cultured cells, cells in suspension, and Gram-negative bacteria. The kit employs a flexible solid-phase system with a unique trifunctional purification matrix and reagents for lysing cells and capturing and releasing DNA. This method does not require prelysing of the sample or precipitation of the DNA.

The Generation Capture Plate Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.
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Cross-contamination-free PCR amplification.

Whole blood or PBS (200 µl) was dispensed into alternate wells of a Generation Capture Plate. DNA was purified using the Generation Capture Plate Kit and PCR amplified using HLA-H locus primers. The 390 bp HLA-H amplicon was detected in all 48 positive controls. No cross-contamination was detected in any of the 48 negative controls.

Performance

DNA yields depend on the number of nucleated cells present. Yields from whole blood may vary widely since white blood cell counts can differ by as much as tenfold. Typical DNA yields obtained from 200 μl whole blood with a normal white blood cell count (5–7 x 106 cells/ml blood) are in the range of 3–8 μg DNA.

The Generation Capture Plate procedure yields pure DNA, indicated by A260/A280 ratios greater than 1.4. Up to 95% of the purified DNA is greater than 23 kb in size. The purified DNA can be stored at 2–8°C for up to 3 months. For long term storage (>3 months) store DNA at –20°C.

Principle

The Generation Capture Plate Kit employs a flexible solid-phase system with a unique trifunctional purification matrix and reagents for lysing cells and capturing and releasing DNA. This method does not require prelysing of the sample or precipitation of the DNA.

Procedure

A sample is applied to the purification matrix contained in a 96-well plate. The cells contained in the sample lyse upon contact with the matrix. Once the cells are lysed, DNA is captured by the matrix material, which makes it possible to efficiently wash away contaminants, leaving the DNA bound to the matrix. Contaminants, including protein, heme, and RNA are removed from the matrix by washing with DNA Purification Solution 1. Following removal of contaminants, the DNA is released from the matrix with DNA Elution Solution 2 and heat. Samples of purified DNA are ready for analysis and do not require precipitation. The purified DNA solution is compatible with PCR and other DNA amplification technologies but is not compatible with real-time PCR, restriction enzyme digests, or Southern blot analysis.

Applications

The purified DNA is compatible with PCR and other DNA amplification technologies but is not compatible with real-time PCR, restriction enzyme digestion, or Southern blot analysis. The performance characteristics of this product have not been fully established.

Features
Specifications
Applications Qualitative PCR
Elution volume 100 - 200 µl
Format Spin column
Main sample type Whole blood
Processing Manual (centrifugation)
Purification of total RNA, miRNA, poly A+ mRNA, DNA or protein Genomic DNA
Sample amount 200 µl
Technology Solid phase with unique trifunctional matrix, no need for separate lysis step
Time per run or per prep 70 min for 96 samples
Yield 3 - 8 µg
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