Bio-Imaging Systems Ethidium Bromide - berthold …

1 Bio-Imaging SystemsParameterValueExposure SecondsGain1 Brightness0 Scan ModeFastIllumination470nmFilterOrange (standard filter)Application NoteEthidium BromideEthidium Bromide is an intercalating agent which resembles aDNA base pair. Due to its unique structure, it can easilyintercalate into DNA strand. Therefore, it is commonly usedas nucleic acid fluorescent tag in various techniques of the lifescience field. The Ethidium Bromide is being in used since the1950', when used in veterinary treat for cattle. It is commonknowledge the EtBr may act as a strong mutagen, while it scarcinogenic and teratogen aspects still have to be well it toxicity, it define as harmful by all routes of entry;inhalation, ingestion, or skin absorption.

2 The EtBr causes eye,skin, mucous membranes and upper respiratory tract to most of the fluorescent compounds, the EtBr is anaromatic molecule. It's core can be defined as phenanthridine an isomer of reason for Ethidium Bromide 's intense fluorescence afterbinding with DNA is the hydrophobic environment foundbetween the base pairs. By moving into this environment andaway from the solvent, the EtBr cation is forced to shed anywater associated molecules. As water is a highly efficientfluorescent quencher, the removal of water molecules allowsthe EtBr to EtBr common dye has several biological DNA it is usually used for double strand from PCR,restriction digest etc.

3 SsRNA can also be detected, since itusually folds back onto itself and consequently provides localbase pairing for the dye Ethidium Bromide can be detected in most kinds of the biological experiment while using its excitation spectrum peaks (300nmand 520nm) and the emission peak at Laboratories Detection TipThe EtBr dye can be easily detected by the MicroBIS with the 470nmexcitation source and the standard orange are several configurations which can be used to detect this dye,and the user must choose the ultimate parameters for his/her experiments,according to his experiment s specification (camera settings andillumination).

4 General parameters for EtBr dye detection in the MicroBIS:General Information:Chemical Name: Ethidium BromideChemical Formula: C21H20N3 BrMolecular Weight: : Ethobromide; Dromilac; Homidium Bromide ; EtBr; RD 1572 and ExcltationWave length (nm)300500600700400 Ethidium Bromide excitation and emissionspectrums (Accordingly in blue and red)Image courtesy of Invitrogen650550450350250 Fluorescence Emission Ethidium Bromide structureEtBr detection by the MicroBISE lectrophoresis was carried out with Low DNA Mass Ladder .DNR EtBr ImageGeneral Information: Bio-Imaging system : MicroBISGel: E-Gel 2% with SYBR Safe , Catalog number: G6000-02, InvitrogenSample: Low DNA Mass Ladder Catalog number 10068-013, , O.

5 Ethidium Bromide and Safety- Readers Suggest Alternative Solutions. Trends Genet. 4:89; , ; Zhang, Y.; Dowling, ; Jin, N.; Adamska, M.; Shiga, K.; Szigeti, K.; Shy, E. S.; Li, J.; Zhang, X.; Lupski, ; & Meisler, Mutation of FIG4 causes neurodegeneration in the pale tremor mouse and patients with CMT4J. Nat. 448:68-72; 2007 Gaw, ; & Patel, HYPERLINK " " Small-moleculeinteraction with a five-guanine-tract G-quadruplex structure from the human MYC promoter Anh Tu n Phan, Vitaly Kuryavyi, Biol. 1:167-173; 2005 Lunn, G.

6 ; Sansone, Decontamination of Ethidium Bromide Spills. Appl. Ind. Hyg. 4: 234-237; , J. III; Kearns, Mechanism of Ethidium Bromide fluorescence enhancement on binding to nucleic acids. Bioche. 16(16):3647 3654; 1977 Application Note | Ethidium BromideLanes above image shows that the MicroBIS minimum detection with EtBr is By using the MicroBISUV light table the minimum detection increase in one fold (down to pg). Bio-Imaging SystemsTel: +972 2 toll-free: 1 866 Technologies for Breakthrough ResultsThis Technical Note is provided as a guideline, and is not intended to bea guarantee of performance.