All samples were analyzed in parallel by digestion with NlaIII, MslI, and BstX, and the fragments were resolved by 2% agarose gel electrophoresis and ethidium bromide
staining (data not shown).
Care is also required using ethidium bromide
. Students must wear gloves.
With careful attention to factors such as the timing of ethidium bromide
staining, background can be reduced but not eliminated, probably owing much to scattered fluorescence from nearby lanes (6).
Assuming 10-20 pg total RNA per thyrocyte, mRNA from ~50-100 cells/mL of blood was detected by ethidium bromide
The PCR products were electrophoresed on 4% agarose gel and stained with ethidium bromide
Amplified PCR products along with the molecular weight marker, 100 bp DNA ladder (GIBCO BRL), were separated electrophoretically (2% agarose gel at 75 V for 60 min) and visualized by ultraviolet illumination after staining with 0.5 [micro]g/mL ethidium bromide
. Gels were photographed with Type 55 positive/negative film (Polaroid, Cambridge, MA).
The DNA was amplified in a 25-[micro]L reaction mixture containing 1.6 [micro]M each of FIP and BIP, 0.2 [micro]M each of the outer primers, 1.6 mM dNTPs, 1 M betaine (Sigma), 20 mM Tris-HCl (pH 8.8), 10 mM KCl, 10 mM (NH4)2S[O.sub.4], 4 MM MgS[O.sub.4], 1.0 mL/L Triton X-100, 8 U of the Bst DNA polymerase large fragment (New England Biolabs), and 0.25 mg/L ethidium bromide
. Denaturation of plasmid DNA (0.2 ng/mL in Tris-EDTA, pH 7.5) was performed at 95[degrees]C for 5 min followed quickly by placement on ice for 5 min.
DNA concentration was assessed by the intensity of the DNA band after agarose gel electrophoresis and staining with ethidium bromide
. Known amounts of [lambda]-DNA were used as a standard.
The fluorescence intensity of ethidium bromide
changes with changes in the oligonucleotide base pairing (as in nuclease digestion) (18).
In this study we used a direct injection of Ethidium bromide
(EB) into the intracerebral ventricle (ICV) as a simple model for induction of neural cells degeneration and MS disease.
PCR was performed and its products were electrophoresed in an ethidium bromide
stained 1% agarose gel with I>>-Hind III digest as a marker 1.