We usually use 6X DNA Loading Dye. We mix 5 uL of DNA with 1 uL of 6X Loading Dye. Final volume is 6 uL, resulting in 1X concentration of the Dye. Hope this makes sense.
Please see the previous comment "We usually use 6X DNA Loading Dye. We mix 5 uL of DNA with 1 uL of 6X Loading Dye. Final volume is 6 uL, resulting in 1X concentration of the Dye. Hope this makes sense." 6x loading dye means it is 6 times too concentrated. You need to dilute it 1/6 so it is 1x loading dye. 1 uL dye + 5 uL DNA = 6 uL total solution. 1 uL dye / 6 uL total solution = 1/6
x = times; therefore 6x loading dye = 6 times loading dye. This meaning it is 6 times more concentrated then the working stock. I is made this way to mix 5 uL of DNA with every 1 uL of 6x loading dye. At this dilution (1/6) it will be 1x loading dye. Hope this helps.
In the old days we used ethidium bromide to stain the DNA and a UV light to cause the ethidium bromide to fluoresce. Both ethidium bromide and UV are somewhat dangerous. We now us a chemical call Red-Safe to stain the DNA. This does not require UV light. Instead we use a blue light and an orange filter to visualized the nucleotide bands.
Thank you so much for your video!
very good explanation!
Thanks a lot for replying
What was the amount and ratio used in
loading dye : DNA
We usually use 6X DNA Loading Dye. We mix 5 uL of DNA with 1 uL of 6X Loading Dye. Final volume is 6 uL, resulting in 1X concentration of the Dye. Hope this makes sense.
How could I do the 6X? And do you recommend use 3 ul of DNA and 7ul of 6X? I have watched it some time...
Please see the previous comment "We usually use 6X DNA Loading Dye. We mix 5 uL of DNA with 1 uL of 6X Loading Dye. Final volume is 6 uL, resulting in 1X concentration of the Dye. Hope this makes sense."
6x loading dye means it is 6 times too concentrated. You need to dilute it 1/6 so it is 1x loading dye.
1 uL dye + 5 uL DNA = 6 uL total solution. 1 uL dye / 6 uL total solution = 1/6
What is x here ?
x = times; therefore 6x loading dye = 6 times loading dye. This meaning it is 6 times more concentrated then the working stock. I is made this way to mix 5 uL of DNA with every 1 uL of 6x loading dye. At this dilution (1/6) it will be 1x loading dye. Hope this helps.
What’s that filter you used to see the DNA bands?
In the old days we used ethidium bromide to stain the DNA and a UV light to cause the ethidium bromide to fluoresce. Both ethidium bromide and UV are somewhat dangerous. We now us a chemical call Red-Safe to stain the DNA. This does not require UV light. Instead we use a blue light and an orange filter to visualized the nucleotide bands.