binding with traditional alkaline lysis of bacterial cells.

he current state-of-the-art methodology in DNA purification relies on the ability of DNA to bind to silica membranes in the presence of high concentration of chaotropic salts such as guanidine. Impurities (e.g. denatured proteins and the attached genomic DNA, RNA, or dNTP and primers in case PCR purification, or dissolved agaorose gel components in case of DNA fragment purification from gel) are then removed by washing with ethanol containing AW buffer, while DNA remains immobilized. Based on this principle, Allele Miniprep Kits and Allele-In-One PCR/Gel Purification Kits provide reliable, convenient, and inexpensive tools for all common DNA purification purposes.

Allele Miniprep Kit, combining DNA-silica

Box 1 | Miniprep Results Catalog Numbers: ABP-PP-MP00500 (50) Preps ABP-PP-MP01000 (100) Preps ABP-PP-MP02500 (250) Preps

Competitors
The image below shows plasmid DNA recovery against competitors. 2 L of each plasmid DNA sample was run on 1% agarose gel for 30 minutes. Miniprep DNA obtained from 3 mL culture of E. coli.
Each batch of reagents is vigorously tested for consistency and stability.

Useful Tips

Description
Allele Miniprep Kits combine DNAsilica binding with traditional alkaline lysis of bacterial cells. The procedure takes aproximately 15 minutes to complete and is suitable for overnight E. coli cultures from 1-6 ml. Recommended for use with recAstrains such as DH5 or XL-1 Blue. Most current commercial plasmid vectors contain high copy number replication origin (e.g. ColE1) and can function well with Allele Miniprep system.

For Allele Miniprep Kits, if you find digestion by certain restriction enzymes is inefficient, there may be carbohydrates overexpressed in certain bacterial strains that remain associated with plasmid DNA with inhibitory effects. To solve the problem of overexpression, wash the cell pellets after initial centrifugation of overnight cultures with 250 ul of STE buffer (TE with 150 mM of NaCl), centrifuge again and proceed to adding buffer A1. For PCR/gel purification, if the DNA is large (>10kb), incubate DNA in column for 5 minutes to allow DNA to bind to membrane more effectively before proceeding to wash with AW buffer. Incubating after addition of TE buffer or H2O may also increase DNA yield.

Box 2 | Contents
Kit Contents Allele Mini-spin Column 2 mL collection tubes (50) 50 50 50 12.5 mL 12.5 mL 17.5 mL 6.25 mL* 5 mL 0.5 mL (100) 100 100 100 25 mL 25 mL 35 mL 12.5 mL* 10 mL 1 mL (250) 250 250 250 62.5 mL 62.5 mL 87.5 mL 31.25 mL * 35 mL 1.5 mL

Allele Miniprep Kit features: Reliabilty: Kits are rigorously tested for quality assurance, so you can expect excellent results every time Convenience: Kits provide everything needed for your experiments, including DNA storage tubes and agarose gel loading dye Reasonable Pricing: Get transfection grade, pure DNA at prices comparable to phenol extraction or boil prep

1.5 mL elution recovery tubes A1: Cell suspension buffer A2: Lysis buffer A3: DNA binding solution AW: Washing buffer TE: TE buffer 10X DNA gel loading dye

*All volumes for AW (washing buffer) indicate volumes before the addition of 95-100% ethanol. (50) reaction/prep kit add 25 mL 95-100% ethanol. (100) reaction/prep kit add 50 mL 95-100% ethanol. (250) reaction/ prep kit add 125 mL 95-100% ethanol

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Protocols
Important Reminders Prior to use, bacteria culture (1-6 mL) must be spun down into pellet. Discard all liquid media in supernatant. Add ethanol to AW buffer (wash buffer). All centrifugation steps at 14k rpm. Procedure 1. Re-suspend pellet using 250 L A1 buffer. Homogenize the mix by vortexing or pipeting up and down. 2. Add 250 L A2 buffer. Mix by inverting tube several times. DO NOT VORTEX. Incubate for one minute. 3. Add 350 L A3 buffer to lysed mixture and invert immediately several times. DO NOT VORTEX. 4. Centrifuge lysed mixture for 10 minutes. 5. Check that Allele Mini-spin column is placed inside 2 mL collection tube. 6. Transfer supernatant from Step 4 into spin column. 7. Centrifuge spin column for 1 minute and discard waste from collection tube. 8. Wash column by adding 600 L AW buffer to column and centrifuge for 1 minute. Discard waste. 9. Centrifuge for 1 minute to remove residual washing buffer. 10. Remove column from collection tube and place into clean microcentrifuge tube. Discard collection tube. 11. Elute DNA by adding 50 L TE buffer directly to column membrane and incubate for 1 minute. 12. Centrifuge for 1 minute. 13. Store DNA at -20C. \

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Storage
Allele Miniprep kits and Allele-In-One PCR/Gel Purification kits should be stored at room temperature in a dry place. Make sure that buffers do not contain precipitates before use. Resuspend precipitates by heating to 35-45C if necessary. All steps are to be carried out at room temperature.

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