DNA Ladepuffer I Fluoro (6x)

Question 1

Is the integrated fluorescent dye non-toxic?

Answer

Yes, the integrated fluorescent dye DNA loading buffer I Fluoro (6x) is a non-toxic nucleic acid dye. It can be easily disposed of with normal waste. Separate handling of the waste as with EtBr is therefore not necessary. If necessary, obtain information from your local waste disposal company.

Question 2

Can I use my existing imaging system for gels with the SimplyEnlight PCR products?

Answer

Yes, you can use UV or blue light for detection of the signal. Therefore, you can replace, e.g., EtBr directly with the DNA loading buffer I Fluoro (6x) without having to change your existing imaging system.

Question 3

Is it necessary to use the GenLadder 100 bp Plus with gel staining dye when using the Red MasterMix Fluoro (2X)?

Answer

If you want to benefit from all the advantages of the SimplyEnlight PCR products, you will also need to use a DNA ladder with fluorescent dye to avoid further staining of the gel. To make it as convenient as possible for you, you can use our GenLadder 100bp Plus with staining dye for this purpose. However, you can take our DNA Ladder 100bp (or any other) and add our DNA Loading Buffer I Fluoro (6X) in a 5:1 ratio before loading the gel.

Question 4

Can I use my own cycling conditions when using the Red MasterMix Fluoro (2X)?

Answer

Yes, you can use your own cycling conditions. Please note that the cycling conditions in our protocol are only guidelines for PCR amplification. Optimal reaction conditions such as incubation times, temperatures, and amount of template DNA may vary and must be determined individually. Besides this, optimization of the annealing temperature is important to guarantee an optimal amplification. We recommend performing a temperature gradient and using primers with a Tm >60°C (Tm = melting temperature of the primer, which is the temperature at which 50% of the primer bind to the complementary sequence of the target DNA.)

Question 5

I have observed a migration shift in the gel electrophoresis using the SimplyEnlight PCR products. What should I do?

Answer

If DNA concentration is less than 4pg, it may cause a migratory shift when performing gel electrophoresis. To remedy this, we recommend removing the fluorescent dye prior to post-staining with our DNA Loading buffer I Fluoro (6x) again for restoring the DNA molecular weight in the original position. To remove the fluorescent dye, immerse the PCR product containing the fluorescent dye into 100mM NaCl and add 2.5 volumes of absolute or 95% ethanol. Incubate on ice for 20 min and centrifuge the mixture at 4°C for at least 10 minutes. Then remove the suspension of ethanol and wash the pellet with 1mL of 70% ethanol. Dry the residual ethanol and resuspend the dsDNA in the TE buffer.

DNA Ladepuffer I Fluoro (6x)

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