Common questions

How do you make 6X loading dye?

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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:

  1. 60% v/v glycerol.
  2. 20 mM Tris-HCL.
  3. 60 mM EDTA.
  4. 0.48% SDS.
  5. 0.03% xylene cyanol.
  6. 0.03% bromophenol blue.
  7. 0.12% Orange G.

What is 6X loading dye?

Thermo Scientific 6X DNA Loading Dye is used to prepare DNA markers and samples for loading on agarose or polyacrylamide gels. It contains two different dyes (bromophenol blue and xylene cyanol FF) for visual tracking of DNA migration during electrophoresis.

How do you make 6X Orange G loading dye?

Composition of 6X Solution 10 mM Tris-HCl (pH 7.6), 0.15% orange G, 60% glycerol, 60 mM EDTA. or 2 ml 50X TAE, 0.15 g Orange G, 60 ml Glycerol fill up to 100 ml with MQ-Water.

How do you make DNA loading dye?

Directions:

  1. Add 25 mg of bromophenol blue to 6.7 ml of ddH2O and mix.
  2. Add 25 mg of xylene cyanol FF and mix.
  3. Add 3.3 ml of glycerol and mix.
  4. Aliquot and freeze at -20 °C for long-term storage.

How many μl of 6X loading dye should you mix with your 10 μl DNA sample before you load the your gel?

2 µl
1. Add 2 µl of UView 6x loading dye to each 10 µl sample of DNA. The final dilution should be 1 part dye to 5 parts DNA sample.

How do I make 6X Laemmli buffer?

Laemmli’s Buffer, 6x

  1. 1.2g SDS (sodium dodecyl sulfate)
  2. 0.01% bromophenol blue.
  3. 4.7ml glycerol.
  4. 1.2ml Tris 0.5M pH6.8.
  5. 2.1ml ddH2O.

What volume is 6X loading dye?

5 µl
Use 5 µl of Gel Loading Dye, Blue (6X) per 25 µl reaction, or 10 µl per 50 µl reaction. Mix well before loading gel.

What is 6X loading buffer?

6X B/Loading Buffer is used as a loading dye for visual tracking of DNA migration during electrophoresis. It incorporates Bromophenol blue. Bromophenol blue migrates fast in the agarose gel and corresponds to the migration of a 300 – 500 bp long DNA fragment in a 1% agarose gel.

What is the composition of gel loading dye?

“A dye used to monitor the migration of DNA into a gel or during gel electrophoresis is known as DNA gel loading dye.” Loading dye is an important component in agarose gel electrophoresis. The loading dye comprises bromophenol blue, Ficoll 400 and water majorly while Xylene cyanol, Tris and EDTA are optional in it.

How much loading dye do I need for gel electrophoresis?

Use 5 µl of Gel Loading Dye, Blue (6X) per 25 µl reaction, or 10 µl per 50 µl reaction. Mix well before loading gel.

How do you make 10X loading dye?

10X Loading Buffer

  1. 2.5 g Ficoll-400.
  2. 1 mL 1M Tris-Cl, pH 7.4. Tris-Cl = Tris-HCl.
  3. 2 mL 0.5 M EDTA.
  4. Add ddH20 to 10 mL, heating to 65oC to dissolve.
  5. Add 25-50 mg of xylene cyanol and 25-50 mg Orange G.

How much loading dye will you add to a 10μl sample before you load it on your gel?

A 10μl sample can be loaded into the agarose gel well. So for 25μl of PCR product roughly add 5 to 7μl of DNA gel loading dye to the PCR tubes. Now you can take 10μl directly from the tubes and load it into the well. Do not forget to mix it properly before loading.

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