products | nucleic acid and protein tools | Nucleic Acid Purification | AquaGenomic™

Nucleic Acid Purification

AquaGenomic™
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A novel aqueous solution-based tenology for isolation and purification of pure genomic DNA

Overview

AquaPlasmid FAQ

1. Q.: Do I need to store the AquaGenomic^TM solution in the refrigerator or freezer?
   A.: No, AquaGenomic^TM Solution is stable at room temperature (~22° C). Invert the bottle a few times to mix the solution well before use.
   
   
2. Q.: Should I keep the solutions and samples on ice while carrying out the isolation?
   A.: No. Most of the isolation steps should be performed at room temperature (~22° C). For DNA isolation from animal cells, cell lysis at 60 °C may increase the yield by ~25%. For DNA isolation from cells with a cell wall, such as bacterial and plant cells, the cell lysis should be carried out at 60-90 °C for 15-30 min.
   
   
3. Q.: Does AquaGenomic^TM Solution contain Proteinase K?
   A.: No. AquaGenomic^TM Solution to extract genomic DNA from cells and tissues can be used without proteinase K digestion. This reduces the time required for genomic DNA isolation from solid tissues from a full day to only ~20 minutes.
   
   
4. Q.: What type of homogenizer do you recommend for using with AquaGenomic^TM ?
   A.: Pestle-and-tube homogenizers produce more homogeneous samples. However, if you do not have a homogenizer, you can chop the tissue with a pair of tweezers, and then vortex it in AquaGenomic^TM Solution to extract the DNA. Another option is to use the 60 °C extraction method, which does not require a homogenizer but a few hours of 60 °C incubation. At the end of incubation, you may use glass or ceramic beads to disrupt the tissue. If beads are not available, a microfuge tube pestle or a pipette tip can be used to chop the tissue.
   
   
5. Q.: Does AquaGenomic^TM Solution contain RNase A?
   A.: No. AquaGenomic^TM Solution can remove most RNA contaminants. Trace amounts of RNA would not interfere with most genomic DNA applications. If complete RNA removal is desired, RNase A (not included; Order No. RIBA25) can be added to AquaGenomic^TM Solution at 25 mg/ml and incubated the lysate at 37-65 °C for 5 min to degrade the RNA.
   
   
6. Q.: Why do I need to purchase a separate reagent for DNA isolation from stool or soil?
   A.: Due to the presence of large amount of enzyme inhibitors in feces and soil, DNA isolation from these samples are particularly challenging. Precipitation of the DNA with isopropanol cannot remove some of the inhibitors and a specially formulated AquaPrecipi Solution (Order No. 3015MT) is required for selective precipitation of the DNA to remove these enzyme inhibitors.
   
   
7. Q.: Can I use AquaGenomic^TM to isolate DNA from frozen blood?
   A.: Yes. The AquaGenomic^TM Blood Protocol uses low concentration of AquaGenomic^TM Solution to lyse RBC (1 volume of AquaGenomic^TM Solution to 20 volumes of whole blood). This RBC lysis method does not depend on a functional RBC membrane and, therefore, can be used for lysis of RBC from either fresh or frozen blood.
   
   
8. Q.: I am concerned about cross-contamination using homogenizers, any tip?
   A.: Yes. Between uses, you should thoroughly wash the homogeniser with soap and running water, soak it in 10% bleach for ~5 minutes, and then rinse it with deionized water. This will prevent crosscontamination of the genomic DNA. If you still feel uneasy, you can use the 60 °C extraction and the bead milling method to disrupt the tissues.
   
   
9. Q.: What genomic DNA yield can I expect using AquaGenomic^TM ?
   A.: Approximately 10 µg DNA can be obtained from 1-2 million cultured cells, 300-400 µl of whole blood, 1 ml of overnight microbial culture, 5-10 mg of animal tissues, or 10-20 mg of plant tissues. DNA yield is dependent on the number of nucleated cells in the sample and may vary at different cell cycles and for different cell types.
   
   
10. Q.: How do I adjust the amount of AquaGenomic^TM Solution based on the amount of starting material?
    A.: Using too much cells/tissues and too little AquaGenomic^TM Solution could result in less clean DNA preparation. As a general guideline, you should use 100 µl of AquaGenomic^TM Solution for 1-2 million cultured cells, 500 µl of whole mammalian blood, 1 ml of overnight microbial cultures, half a buccal swab, or 5 mg of animal or plant tissues.
    
    
11. Q.: How pure is the genomic DNA isolated by AquaGenomic^TM ?
    A.: Typical A260/A280 of AquaGenomicTM purified DNA is 1.6-1.8. The isolated genomic DNA is free from most cellular contaminants and enzyme inhibitors. However, there may be some RNA
    contamination, if RNase A treatment is not included.
    
    
12. Q.: What QC tests do you use to certify your products?
    A.: Each lot is tested to ensure its performance and reliability in isolating genomic DNA. The isolated DNA (a) should have an A260/A280 > 1.6, and (b) should be readily digested with restriction enzymes.

See also chapter 6 of our online catalog.

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