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.
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.
Thank you so much for your video!
Thanks a lot for replying
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.
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.