Product overview Technical/Spec Support/Documents FAQsApplication references Related products DescriptionFluoroDye™ DNA Fluorescent Loading Dye is a ready-to-use 6X DNA loading dye designed for fast qualitative electrophoresis analysis. Containing sensitive fluorescent dye with high specific affinity towards double stranded DNA (dsDNA), the FluoroDye™ Fluorescent DNA Loading Dye has negligible background and renders destaining process unnecessary. The FluoroDye™ DNA Fluorescent Loading Dye allows the user to immediately visualize electrophoresis result upon completion or to monitor the electrophoresis in real time. FluoroDye™ DNA Fluorescent Loading Dye is compatible with both the conventional UV gel-illuminating system as well as the less harmful long wavelength blue light illumination system. FluoroDye™ emission as bound to dsDNA is 522 nm, while its excitation peaks are at 270, 370 and 497 nm.Features:Excellent for premix with DNA samplesSensitivity: 0.14 ng (DNA)A safer alternative to EtBrCompatibility: suitable to blue or UV lightIncreased cloning efficiency (blue light)Composition FluoroDye™ DNA Fluorescent Loading Dye is stored in 6X concentration in 60% glycerol and buffered with Tris-HCl and EDTA, containing Bromophenol blue, Xylene cyanol FF and Orange G as tracking dyes.StorageProtected from light -20°C for 24 monthsSensitivity of FluoroDye™FluoroDye™ DNA Fluorescent Loading Dye (DL5000) shows a green-yellow fluorescence under blue light excitation. The sensitivity of NS1000 is about 0.14 ng (arrow) for a 4 kb fragment. Excitation and emission spectrum of FluoroDye™ FluoroDye™ DNA Fluorescent Loading Dye (DL5000) emission as bound to dsDNA is 522 nm while its excitation peaks are at 270, 370 and 497 nm.  Contents ComponentVolumeCat. No. FluoroDye™ DNA FluorescentLoading Dye (Green, 6X)1 mlDL5000 FluoroDye™ DNA FluorescentLoading Dye (Green, 6X)5 x 1 mlDL5001  StorageProtected from light -20°C for 24 months ManualManual_DL5000_FluoroDye™ DNA Fluorescent Loading DyeSDSSDS_DL5000 Will FluoroDye™ DNA Fluorescent Loading Dye (DL5000) affect the DNA samples in subsequent experiments?  Using FluoroDye™ DNA Fluorescent Loading Dye (DL5000) does not affect subsequent operations. This is because the fluorescent dye can easily be removed by regular alcohol precipitation or gel elution kits.  Will the FluoroDye™ DNA Fluorescent Loading Dye (DL5000) affect cloning efficiency? Using the fluorescent dye coupled with blue light, the cloning efficiency is increased by about 100 times compared to using EtBr and UV light. Can the  FluoroDye™ DNA Fluorescent Loading Dye (DL5000) be removed from DNA? How can we do it?The staining dye can be removed from DNA with traditional ethanol precipitation, PCR clean up kits, or the gel extraction kits. The ethanol precipitation method can follow conventional molecular cloning or follow the listed protocol. Ethanol precipitation.A.Measure the volume of the DNA sample. B.Add 1/10 volume of 3M sodium acetate, pH 5.2,(final concentration of 0.3 M) - The pH value of 3M sodium acetate must be adjusted with acetate not with HCl. C.Mix well. D.Add 2 to 2.5 volumes of cold 100% ethanol (calculated after salt addition). E.Mix well. F.Place on ice or at -20 °C for >20 minutes.G.Spin at maximum speed in a microfuge for 10-15 min.H.Carefully decant supernatant. I.Add 1 mL 70% ethanol. Rinse and spin briefly. Carefully decant supernatant.J.Air dry or briefly vacuum dry pellet. K.Re-suspend pellet in the appropriate volume of TE, Tris buffer or water.Can the FluoroDye™ DNA Fluorescent Loading Dye (DL5000) stained DNA be used for enzymatic reaction like ligation, enzyme digestion, PCR and so on? Do you have any recommended methods like pretreatment? After removing the fluorescent staining dye from DNA, the DNA can be used for ligation, restriction enzyme digestion or PCR reaction. It is imperative that the DNA is maintained in good quality for bioassay after removing the stained dye. We recommend removing the fluorescent staining dye with the gel extraction kits or PCR clean up kits because these methods are convenient and high efficiency. The very low amount of fluorescent staining dye does not affect ligation, enzyme digestion or PCR. However, the threshold is related to the enzyme systems and it is on a case-by-case basis. For the best results, we recommend to remove the fluorescent staining dye from DNA before proceeding to the next step of the experiment. As the FluoroDye™ DNA Fluorescent Loading Dye (DL5000)  is used, is the migration of DNA affected by the fluorescent dye?Yes, it might be affected due to variation of DNA amount mixed with the fluorescent loading dye. The fluorescence composition in a fluorescent loading dye is fixed, so for different amounts of DNA different degrees of binding to fluorescent dyes are expected. Variety in DNA/fluorescence ratio might lead to variation in DNA migration; the higher the fluorescence ratio is, the slower the DNA fragments migrate. In general, a DNA amount of ≥31.25 ng with 1 μL DL5000 loading dye will be an acceptable range. If the DNA concentration is lower than 31.25 ng, the molecular weight determination may be slightly distorted. For analyzing DNA sample of unknown concentration, it is recommended to use the FluoroStain™ DNA staining dye with post-staining method or to use FluoroVue™ nucleic acid staining dye with in-gel staining method.   FUS interacts with nuclear matrix-associated protein SAFB1 as well as Matrin3 to regulate splicing and ligand-mediated transcriptionAtsushi Yamaguchi, Keisuke Takanashi Sci Rep. 2016; 6: 35195. Published online 2016 Oct 12. doi: 10.1038/srep35195PMCID: PMC5059712PCR analysis for assessment of bacterial bioburden in orthokeratology lens casesJung Lo, Po-Chiung Fang, Chun-Chih Chien, Chang-Chun Hsiao, Shin-Ling Tseng, Yu-Hsuan Lai, Ming-Tse Kuo Mol Vis. 2016; 22: 1–8. Published online 2016 Jan 14. PMCID: PMC4734148Carriage rates of Neisseria meningitidis serogroups: determination among freshmen conscripts before vaccinationRamezan Ali Ataee, Ali Mehrabi-Tavana, Seyed Mohammad Javad Hosseini, Farshad Kaviani Iran J Microbiol. 2016 Jun; 8(3): 203–209.PMCID: PMC5139924Helicobacter pylori neutrophil-activating protein induces release of histamine and interleukin-6 through G protein-mediated MAPKs and PI3K/Akt pathways in HMC-1 cellsChung-Che Tsai, Ting-Yu Kuo, Zhi-Wei Hong, Ying-Chieh Yeh, Kuo-Shun Shih, Shin-Yi Du, Hua-Wen Fu Virulence. 2015 Nov-Dec; 6(8): 755–765. Published online 2015 Sep 16. doi: 10.1080/21505594.2015.1043505 PMCID: PMC4826132 B-BOX™ Blue Light LED Epi-illuminator470 nm long wavelengthImproved cloning efficiency Compact, light-weight, and portable (less than 1 kg)Adjustable and removable filter plate allows for gel cutting, visualization, and documentationFluoroBand™ DNA Ladder seriesSharp bandsQuick reference— enhanced bands Ready-to-use— premixed with loading dye for direct loadingStable— room temperature storage over 6 monthsDirectly observed by UV or blue light— premixed with high sensitive fluorescent dyeFluoroVue™ Nucleic Acid Gel StainExcellent for in-gel stainingSensitivity up to 0.14 ng (DNA) or or 1 ng (total RNA)A safe alternative to EtBrSuitable for blue or UV light