FAQ about AquaGenomic
1. Do I need to keep AquaGenomic in the freezer?
No, AquaGenomic Solution is stable at room temperature (~22 °C) for >1 year.
2. Does AquaGenomic Solution contain Proteinase K?
No. AquaGenomic can be used to extract DNA from most cells and tissues without needing protease digestion. However, adding Proteinase K (50 mg/ml) to AquaGenomic solution can increase DNA yield and is required for mitochondrial DNA extraction. You may homogenize the sample in AquaGenomic containing Proteinase K, incubate it at 55 °C for 1-2 hrs and then at 95 °C for 10-15 min to inactivate the Proteinase K.
3. I am worried about cross-contamination using homogenizers, any tips?
Between uses, you may wash the homogenizer with soap and running water, soak it in 10% bleach for ~5 min, and then rinse it with running deionized water. If you still feel uneasy, you may use Proteinase K digestion without using a homogenizer, or use a multichannel bead beater for homogenization in screw-capped tubes.
4. How may I maximize the DNA yield?
Incubating the DNA pellet in TE buffer or deionized water at 22 or 4 °C overnight before centrifuging to remove the insoluble may maximize DNA recovery.
5. Do I have to use the lysate immediately for PCR?
No, you may store the lysate at 4 °C until analysis. If the lysate has been incubated at 75-85 °C for 20 min, it may even be left at room temperature until analysis.
6. I got a weak PCR amplification using the lysate directly, how may I improve it?
You may try a few things to optimize the amplification: a) try to use different amount of lysate for the PCR, from 0.25 ml undiluted lysate to 20x diluted lysate, b) add 0.1 mg/ml BSA to the PCR reaction, c) add 1 mg/ml DTT to the PCR reaction, and d) increase the PCR cycle number to 45 cycles.