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QuickExtractTM

DNA Extraction Kit

QuickExtract™ DNA Extraction Solution

Applications
  • Simple, rapid extraction of PCR-ready DNA for transgenic mouse genotyping, genetic studies, human identity testing, or viral/microbial screening.

The QuickExtract™ DNA Extraction Solution can be used to rapidly and efficiently extract PCR-ready genomic DNA from almost any sample type using a simple, one-tube protocol that takes only 3-8 minutes (Fig. 1), depending on the sample. QuickExtract Solution has been used to extract DNA from samples such as hair follicles, quill-end cells of feathers, tissue-culture cells, buccal cells, zebrafish organs and scales, and mouse tail snips. The extracted DNA is suitable for PCR analysis (Fig. 2), such as genomic, transgenic, or viral DNA screening in animals, or for genetic or environmental research and screening in humans and other organisms.

The QuickExtract method allows for the inexpensive processing of one to hundreds of samples simultaneously, without centrifugation, spin columns or the use of any toxic organic solvent. The method is also compatible with robotic automation.

Benefits
  • Nontoxic reagents, inexpensive processing.
  • Short procedure (8 minutes for the average sample).
  • No centrifugation or spin columns to reduce yields.
  • Compatible with high-throughput and robotic workflows.


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Figure 1. Procedure for obtaining PCR-ready DNA using QuickExtract™ DNA Extraction Solution.

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 Figure 2. FailSafe™ PCR amplifications of genomic DNA extracted from a variety of tissues or cells. Buccal cells were extracted using the BuccalAmp™ DNA Extraction Kit, and all other samples with QuickExtract™ DNA Extraction Solution. PCR was performed using primers to amplify the regions indicated: Lanes 1-3, human β-globin; lane 4, transgenic mouse GAPDH; lane 5, E. coli 16S ribosomal RNA gene; lane 6, transgenic SV40 T antigen. Figure 3. Extracted DNA from multiple Zebrafish organs using QuickExtract™ DNA Extraction Solution 1.0. A 1-µl aliquot of a 100-µl extracted sample was used to amplify a single-copy crystallin-like gene. Lane 1, 100-bp ladder; lanes 2-3, fins; lanes 4-5, eyes; lanes 6-7, scales; lane 8, no-DNA control. 





QuickExtract™ Bacterial DNA Extraction Kit

Applications
  • Preparation of DNA from Gram-positive and Gram-negative bacteria.
  • DNA can be used for PCR, restriction digests, pulsed-field gel electrophoresis (PFGE), and optical mapping.

The QuickExtract™ Bacterial DNA Extraction Kit provides a simple method for extracting DNA from Gram-positive and Gram-negative bacteria. To a washed pellet of the bacterial sample, add QuickExtract Bacterial DNA Extraction Solution and Ready-Lyse™ Lysozyme Solution. Ready-Lyse Lysozyme is a proven reagent with over 200 times the specific activity of egg-white lysozyme. Mix and incubate for 15 minutes at room temperature. For some species, if lysis is not achieved, the incubation time can be extended. Samples may optionally be heated at 80°C for 2 minutes to kill any remaining viable bacteria. The DNA is now ready for PCR or other downstream applications such as restriction digests, PFGE (pulsed-field gel electrophoresis), and optical mapping. The kit has been tested on a range of bacteria (Table 1) and has been shown to produce very long DNA strands (Fig. 1).


Benefits
  • Single-tube system.
  • No toxic organic solvents.
  • Suitable for high-throughput applications.
  • Ready-Lyse™ Lysozyme supplied.
  • Long DNA generated.

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Table 1. Species of bacteria that have been tested with the QuickExtract™ Bacterial DNA Extraction Kit.
 
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 Figure 1. Long DNA strands from E. coli extracted using the QuickExtract™ Bacterial DNA Extraction Kit. Many strands are 400-500 kb. Image courtesy of Dr. Shiguo Zhou, University of Wisconsin-Madison.










QuickExtract™ Plant DNA Extraction Solution

Applications
  • High-throughput isolation of DNA from plant leaf samples for PCR-based analysis, e.g., GMO testing.

The QuickExtract™ Plant DNA Extraction Solution can be used to rapidly and efficiently extract PCR-ready genomic DNA from most plant leaf samples using a simple, one-tube protocol that takes only 8 minutes (Fig. 1). Most leafy plants are suitable for DNA extraction using the QuickExtract Plant Solution, including Arabidopsis, barley, maize, emmer, pepper, rice, spelt, spinach, soybeans, and wheat (e.g., Fig. 2).


The QuickExtract Plant method allows for the inexpensive processing of one to hundreds of samples simultaneously, without grinding the sample, centrifugation, spin columns, or any toxic organic solvent. The procedure is fully compatible with robotic automation, provides a PCR-ready sample, and is reproducible (Fig. 3). Simply add the QuickExtract Plant solution to the sample and perform two sequential heating steps. A small aliquot of the sample is then used as a template for PCR or qPCR.

Benefits
  • qPCR or PCR-ready sample.
  • No bead-beating, freezing, or grinding of plant leaf material.
  • Nontoxic, inexpensive processing.
  • Fast procedure (8 minutes for average sample).
  • No centrifugation or spin columns to reduce yields.
  • Suitable for high-throughput or automated workflows.

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Figure 1. Overview of the QuickExtract™ Plant DNA extraction procedure.

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 Figure 2. PCR products using QuickExtract™ Plant DNA Extraction Solution with different varieties of plant leaves. Forty cycles of RAPD were performed with DNA extracted from plant leaves using the QuickExtract Plant solution and the FailSafe™ PCR System. Lane M, 100-bp ladder; lane 1, pepper; lane 2, soybean; lane 3, spelt.

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 Figure 3. Reproducibility of PCR results with QuickExtract™ Plant DNA Extraction Solution from Arabidopsis thaliana leaves.Six individual punches of four Arabidopsis leaves were treated with QuickExtract Plant solution as described in the product information sheet. One microliter of the solution was used in a 25-µl PCR using the FailSafe™ PCR System and primers specifi c for the single-copy HSC70 chromosomal gene. Aliquots were analyzed on a 2% agarose gel and the DNA was visualized by staining with SYBR® Gold. Lanes 1-6, PCR products of extracted DNA samples; lane 7, no-leaf negative control; lane M, 100-bp DNA ladder. The expected amplicon is approximately 520 bp.



QuickExtract™ Seed DNA Extraction Solution

Poster from Plant and Animal Genome Conference 2009 (637k)

Applications
  • High-throughput isolation of DNA from ground seed samples for PCR-based analysis, e.g., GMO testing.

The QuickExtract™ Seed DNA Extraction Solution can be used to rapidly and efficiently extract PCR-ready genomic DNA from most ground seed samples using a simple, one-tube protocol that takes only 8 minutes (Fig. 1). The method allows for the inexpensive processing of one to hundreds of samples simultaneously, without centrifugation, spin columns or use of any toxic organic solvent


The procedure is fully compatible with robotic automation, provides a PCR-ready sample, and is reproducible (Fig. 2). Simply add the QuickExtract Seed solution to the ground sample and perform two sequential heating steps. A small aliquot of the sample mix is then used as a template for PCR or qPCR. Epicentre recommends PCR optimization with the FailSafe™ PCR PreMix Selection Kit or a fast PCR with the TAQXpedite PCR System. Seeds tested include apple, barley, cotton, maize, oat, rice, rye, sunflower, switchgrass, tomato, and wheat.

Note: Most ground seed material under 10 mg is acceptable for DNA extraction. Extracted DNA from larger sample sizes will not produce satisfactory PCR results.

Benefits
  • Nontoxic reagents, inexpensive processing.
  • Short procedure (8 minutes for the average sample).
  • No centrifugation or spin columns to reduce yields.
  • Suitable for high-throughput or automated workflows.

"Your QuickExtract Seed and PlantAmp helped me resurrect some DNA templates that I was on the verge of abandoning due to lack of quality DNA and subsequent PCR amplification for downstream sequencing."


Millie Burrell
Texas A&M University

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Figure 1. Overview of the QuickExtract™ Seed DNA extraction procedure.

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 Figure 2. PCR of DNA extracted from brown rice seeds. Samples (10 mg each) of five different brown rice seeds were used for DNA extraction, and a 1-µl aliquot of each was used for PCR of the single-copy RIA gene. Lane M, 100-bp ladder; lane 1, no-template control; lanes 2-6, PCR products from five different seed samples.



QuickExtract™ FFPE DNA Extraction Kit

Applications
  • Isolation of DNA from FFPE samples for PCR-based analysis, e.g., microsatellite detection, SNP detection, tumor heterogeneity studies, copy number detection, methylation analysis, and Short Tandem Repeat (STR) analysis.

The analysis of nucleic acid from formalin-fixed, paraffin-embedded (FFPE) specimens is challenging due to the extensive cross-linking of all tissue components during the fixation process. These challenges include chemical modification of the DNA, cross-linking of DNA with other molecules, degradation of the DNA, and the limited amount of nucleic acid in the samples.


The QuickExtract™ FFPE DNA Extraction Kit is a fast, simple, and inexpensive method for preparing genomic DNA for PCR amplification from archival samples (Fig. 1). The protocol requires only heat treatment to melt the paraffin, lyse the cells, decrease the fomalin-induced cross-linking in the sample, and degrade compounds inhibitory to amplification. Following heat treatment, the sample DNA is ready for PCR (Figs. 2 and 3).

Note: This product is only intended for PCR- and qPCR-based applications. If DNA is required for other molecular biology applications, consider the MasterPure™ DNA Purification Kit.

Benefits
  • PCR-ready DNA in minutes, not days.
  • No xylene or phenol extraction.
  • No columns, transfers, or sample loss.
  • Extracted DNA is compatible with both real-time and endpoint PCR.

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Figure 1. Overview of the QuickExtract™ FFPE DNA extraction procedure.

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 Figure 2. PCR amplification of DNA from a slide-mounted, FFPE preserved human skeletal muscle tissue section, extracted with the QuickExtract™ FFPE DNA Extraction Kit. Two microliters of undiluted, extracted DNA was amplified with primers for three different loci: tumor protein 53 (TP53), dystrophin (DMD), and tumor necrosis factor (TNF). The products were separated on a 3% agarose gel and were visualized with SYBR® Gold. Lane M, 100-bp DNA ladder; lane 1, exon 2 of TP53; lane 2, exon 3 of TP53; lane 3, exon 11 of TP53; lane 4, exon 6 of DMD; lane 5, exon 50 of DMD; lane 6, exon 3 of DMD; lane 7, exon 4 of TNF. Figure 3. PCR amplification of DNA from slide-mounted, FFPE-preserved, human tissue sections, combined and extracted with the QuickExtract™ FFPE DNA Extraction Kit. Two microliters of undiluted, extracted DNA was amplified with primers for seven different STR loci. The products were separated on a 3% agarose gel and were visualized with SYBR® Gold. The expected allele-to-allele variation in STR sequences results in multiple products per primer set. Lane M, 100-bp DNA ladder; lane 1, RENA4; lane 2, D3S1358; lane 3, D7S820; lane 4, THO1; lane 5, D19S253; lane 6, D21S11; lane 7, AMEL. 




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