What is the function of Xylene cyanol?
Xylene cyanol can be used as an electrophoretic color marker, or tracking dye, to monitor the process of agarose gel electrophoresis and polyacrylamide gel electrophoresis. Bromophenol blue and orange G can also be used for this purpose.
Table of Contents
What is the function of Xylene cyanol?
Xylene cyanol can be used as an electrophoretic color marker, or tracking dye, to monitor the process of agarose gel electrophoresis and polyacrylamide gel electrophoresis. Bromophenol blue and orange G can also be used for this purpose.
What colour is Xylene cyanol?
What color is xylene Cyanol? Composition: Water 99.85%, Xylene Cyanol FF 0.10%, Methyl Orange, Sodium Salt 0.05% Boiling Point: Approximately 100°C Density: 1 Melting Point: 0°C Color: Dark blue-green liquid Physical State: Liquid pH Range: 2.9 (purple) – 4.6 (green) Solubility Information: Miscible Shelf Life:…
How does xylene Cyanol migrate in the agarose gel?
In agarose gels, Bromophenol Blue and Xylene Cyanol will migrate at approximately 3000 and 300 bp, respectively. These dyes will migrate at different rates in acrylamide gels depending on the gel density. Table 2 provides the approximate migration rate in terms of the relative size of single-stranded/denatured DNA.
What size does xylene Cyanol run at?
To answer your question, bromophenol will run at ~25 nt (nucleotides) and xylene cyanol 100-110 nt (although it depends whether you’re running DNA or RNA as equivalent length molecules run differently owing to the greater mass of RNA).
What causes fuzzy bands in gel electrophoresis?
Primer-dimers The bright fuzzy bands at the bottom of the gel are typical of those caused by primer dimers. Primer-dimers can be minimized by using hot start PCR or by using a higher annealing temperature.
How do you make 6x loading dye?
With 6x dye, load equivalent ratio of 5 µL dye to 25 µL sample. Recipe 1: 0.25 g bromophenol blue. 3 mL glycerol….Recipe 3:
- 60% v/v glycerol.
- 20 mM Tris-HCL.
- 60 mM EDTA.
- 0.48% SDS.
- 0.03% xylene cyanol.
- 0.03% bromophenol blue.
- 0.12% Orange G.
How do you make gel loading dye?
Directions:
- Add 25 mg of bromophenol blue to 6.7 ml of ddH2O and mix.
- Add 25 mg of xylene cyanol FF and mix.
- Add 3.3 ml of glycerol and mix.
- Aliquot and freeze at -20 °C for long-term storage.
Why do gels smear?
If the gel is not poured correctly, it will not polymerize or solidify evenly, thus causing the molecules to smear. If the wells are filled too much, or if the sample is not properly diluted, the excess sample may smear across the gel.
What causes fuzzy bands PCR?
Primer-dimers The bright fuzzy bands at the bottom of the gel are typical of those caused by primer dimers. Primer-dimers can be minimized by using hot start PCR or by using a higher annealing temperature. Preventative measures include designing primers that do not form homo- or hetero-dimers.
What would be the result of running the gel too long?
What would happen if the gel was run for too long? The sample bands would move too far and leave the bottom of the gel.
How many μl of 6x loading dye should you mix with your 10 μl DNA sample before you load the your gel?
2 µl
Add 2 µl of UView 6x loading dye to each 10 µl sample of DNA.
How do you make 6x gel loading dye?
