EnVISION DNA 染料,可作上样缓冲液,含1条指示带，英文名称：EZ-Vision One™ DNA DYE AS LOADING BUFFER SAMPLE，级别：生物技术级，cas：N/A，保存温度：COLD，详细说明：http://www.amresco-inc.com/media.acux?path=/media/products/coa/COA-N472.pdf EZ-Vision™ One, DNA Dye as Loading Buffer, 6X Supplied in Loading Buffer, 6X containing 15% Ficoll Includes a single tracking dye migrating at 10 bp General Information: EZ-Vision™ is a non-mutagenic fluorescent reagent that produces instant visualization of DNA bands upon UV illumination of agarose gels. Supplied in AMRESCO’s 6X DNA Loading Buffer, EZ-Vision™ forms a tight complex with the sample DNA and co-migrates with it during electrophoresis. DNA bands can be visualized immediately after the run by placing the gel on a standard UV transilluminator so that post-run staining and destaining is completely eliminated. EZ-Vision™ is non-mutagenic and has no hazardous shipping, handling or disposal costs. It is ideal for applications needing rapid DNA band visualization and for environments requiring a safe, non-hazardous alternative to Ethidium Bromide. EZ-Vision™ is available in 2 versions that differ only by the tracking dyes included in the loading buffer. EZ-Vision™ Three contains 3 tracking dyes that migrate at 4,000 bp, 600 bp, and 10 bp. EZ-Vision™ One contains only a single fast-running tracking dye that migrates at approximately 10 bp in a 1% agarose gel. Storage/Stability: EZ-Vision™ is stable for at least 6 months at 2 - 8°C. EZ-Vision™ is light sensitive and should be stored protected from light. Normal usage can be carried out under ambient light. Spectral Information: Excitation = 364 nm Emission = 454 nm Application Disclaimer For Research Use Only. Not for Therapeutic or Diagnostic Use. Protocol: No mutagenic or genotoxic effects are observed in the AMES test or sister-chromatid exchange assay of the EZ-Vision™ reagent. Although it is not toxic at the concentrations used, standard handling precautions are advised for all nucleic acid binding reagents. All local regulations should be followed when using and disposing of this reagent. EZ-Vision™ tracking dyes: Page 2 of 3 EZ-Vision™ Type Color Mobility (1% Agarose) EZ-Vision™Three Light blue 4000 bp Dark blue ~600 bp Magenta ~ 10 bp EZ-Vision™One Magenta ~ 10 bp 1. Vortex EZ-Vision™ for 30 seconds prior to use. 2. Dilute 1 part EZ-Vision™ with 5 parts DNA sample and mix. Note: EZ-Vision™ must be added to DNA markers in order to visualize the ladder bands simultaneously with the sample after electrophoresis. 3. Load sample and run according to standard procedure. 4. After the run, remove gel and place on UV transilluminator to immediately visualize bands. DNA bands will emit blue against a dark background. Fluorescence should be visible on a transilluminator for at least 24 hours after electrophoresis if the gel fluorescence has not been bleached. 5. Gels can be post stained with Ethidium Bromide if desired. Gel Documentation • Black and White Polaroid Photography: EZ-Vision™ stained gels can be photographed using a standard UV transilluminator and Polaroid #667 film with filters such as those used to photograph green dyes such as SyBR™ Green. A filter that is typically used to photograph Ethidium Bromide (EtBr) stained gels can be used, however, EZ-Vision™ dye emits at a much shorter wavelength than Ethidium Bromide. Thus, the EtBr filter is not optimal for EZ-Vision™. Consequently, using a Polaroid camera and standard EtBr filter may require a longer exposure time than if the gel were stained with Ethidium Bromide. Approximately 2 times this exposure time should produce an image for documentation with good contrast which appears to represent the image that you visually perceive. • Gel Imaging Systems: EZ-Vision™ stained gels are compatible with digital imaging systems. Please contact your system manufacturer with the excitation and emission information listed below to obtain information on appropriate filters. Excitation = 364 nm Emission = 454 nm Downstream Applications DNA stained with the EZ-Vision™ probe is compatible with a variety of downstream applications including ligation reactions, transformation procedures and PCR amplification. Results Summary Ligation, transformation and PCR amplification procedures were tested on parallel samples of plasmid DNA fragments stained with either EZ-Vision™ or EtBr. • Recovery from gel slices: The EZ-Vision™ reagent does not interfere with recovery of DNA fragments from agarose gels. Comparison studies from 1% agarose gels were performed to determine the recovery of EZ-Vision™ stained DNA fragments versus EtBr stained fragments. In these tests, a DNA aliquot was combined with EZ-Vision™ and applied to a 1% TAE agarose gel. A second aliquot was loaded on a 1% TAE agarose gel containing 50 μg/ml EtBr. Gels were run according to standard procedures. An 850 bp DNA band was excised from each gel and purified on QIAquick® Gel Extraction Kits (Qiagen Group, Hilden, Germany). Equal aliquots were applied to 1% TAE agarose gels. DNA recoveries were determined by band intensity after electrophoresis. There was no significant difference in the amount of DNA recovered from each gel. • Ligation and Transformation Efficiency: Ligation and transformation efficiency of the EZ-Vision™ stained 850 bp fragment is similar to the EtBr stained fragment. In parallel reactions, the purified 850 bp fragments were ligated into an Ampicillin resistant vector and transformed into E.coli. The transformed cultures were plated onto LB-Ampicillin media. The total number of colonies for each sample was determined after overnight incubation. The number of Ampicillin resistant colonies was similar for each sample. • PCR Amplification: Amplification of an EZ-Vision™ stained DNA fragment was equivalent to the EtBr stained fragment. A 1.3 kb DNA fragment stained with either EZ-Vision™ or EtBr was amplified by PCR using Pfu polymerase. The products were applied to a 1% TAE agarose gel. The 1.3 kb band was excised and purified as described above (see Recovery from gel slices). The recovered DNA samples were quantitated by a NanoDrop® Spectrophotometer (NanoDrop® Technologies, Wilmington, DE). The amount of DNA recovered from each sample was equivalent indicating that EZ-Vision™ stained DNA was a suitable template for PCR reactions. • Sequencing: The use of EZ-VisionTM stained DNA as a sequencing template is currently being evaluated. Related Products Page 3 of 3 Code Product Agarose 0710-500G Agarose I™, 500 g General Use (also available as tablets, K857-100TABS) J234-250G Agarose SFR™ Super Fine Resolution for superior resolution of nucleic acid fragments between 200 and 1,000 base pairs. E776-100G Agarose 3:1 HRB™ High Resolution Blend for resolution of nucleic acid fragments below 1,000 base pairs. Buffers 0658-4L TBE Buffer, 10X Liquid Concentrate 0478-2PK TBE Buffer, 10X Ready-Pack™ 0796-1.6L TAE Buffer, 25X Liquid Concentrate Markers K180-250UL 100 bp Ladder 13 bands ranging from 100-3000 base pairs E854-50RXN PCR DNA Marker™ 8 bands ranging from 50 to 2000 base pairs K181-500UL 1 kb Ladder 11 bands ranging from 500 to10,000 base pairs
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