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Invitrogen™ DNAzol™ Reagent, for isolation of genomic DNA from solid and liquid samples

Catalog No. 10503027
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10503027 100 mL
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Catalog No. 10503027 Supplier Invitrogen™ Supplier No. 10503027
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Description

Includes

1 bottle (100mL)

DNAzol Reagent is a complete and ready-to-use organic reagent from animal, plant, yeast, and bacterial origin.

DNAzol Reagent is a complete and ready-to-use organic reagent from animal, plant, yeast, and bacterial origin. The DNAzol Reagent procedure can be completed in typically 10–30 minutes with DNA recoveries of 70–100%.

  • Versatile: Can use with broad spectrum of sample materials
  • Rapid isolation and high recovery of genomic DNA
  • Efficient isolation of genomic DNA from a variety of sample types
  • Rapid isolation and high recovery of genomic DNA
  • Isolated DNA can be used for a number of downstream applications

Order Info

Shipping Condition: Room Temperature

Specifications

Content And Storage 100 mL DNAzol Reagent; room temperature
Isolation Technology Organic Extraction
Sample Type Bacteria, Cells, Plant, Tissue, Yeast
Elution Volume Varies
Final Product Type Genomic DNA
For Use With (Application) PCR, southern blotting, sequencing, nucleic acid labeling, hybridization
High-throughput Compatibility Not High-throughput Compatible (Manual)
Quantity 100 mL
Shipping Condition Room Temperature
Test Time 30 min.
Starting Material Amount Bacteria: ≤107 cells or 0.1 mL
Cells: ≤107
Plant: ≤50 mg
Tissue: ≤50 mg
Yeast: ≤107 cells or 0.1 mL
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Frequently Asked Questions (FAQs)

I see two phases after addition of ethanol during DNA isolation using DNAzol Reagent. What should I do?

It is possible to see two phases after addition of ethanol if the amount of DNAzol Reagent was too low. Add more DNAzol Reagent and continue.
I've isolated my DNA using the DNAzol Reagent, but cannot cut it with restriction enzymes. What could be causing this?

If the cells or tissue were washed with phosphate buffer solutions prior to DNA isolation, the phosphate may have been carried over and be inhibiting restriction enzymes. We recommend adding DNAzol Reagent to the DNA solution and reprecipitating with 0.5 volumes of 95% EtOH. Wash twice with 95%, dry briefly, and resuspend in 8 mM NaOH.
The DNA I've isolated using the DNAzol Reagent is hard to resuspend. Do you have recommendations for fixing this?

You can try incubating samples resuspended in 8 mM NaOH at 37 degrees C overnight to resuspend the DNA. You can also try incubating at 45 degrees C for 15 minutes.
My DNAzol Reagent turned a dark murky green from its original light green. Is it still okay to use?

Yes, this happens due to the dye in the reagent, and seems to be dependent on the volume of the stored reagent. The color change does not affect its performance.
Will DNAzol Reagent isolate only genomic DNA or will plasmid DNA also be isolated? How about mitochondrial DNA?

The DNA isolated is actually total DNA, so plasmid DNA will be isolated along with genomic DNA. The mitochondrial genome is similar to a plasmid and can be isolated using DNAzol Reagent. The 1 minute room temperature incubation in ethanol before centrifugation should be extended to 5-10 minutes for maximum recovery.
At what temperatures can DNAzol reagent be stored and still be okay to use? Are there possible stopping/storage points in the protocol?

We recommend storing DNAzol Reagent at room temperature. The DNAzol lysate (homogenate) can be stored 1 month at 15-30 degrees C; after 10 months at 4 degrees C or -20 degrees C, the DNAzol lysate (homogenate) has yielded high molecular weight genomic DNA, which can be completely digested with restriction enzymes and works well in PCR. During washes, DNA can be stored in 95% EtOH for at least one week at 15 degrees C to 30 degrees C or for three months at approximately 4 degrees C. DNA can be stored in DNAzol Reagent for one month at room temperature or 10 months at 4 degrees C.
For genomic DNA isolation, should I trypsinize my cells before adding the DNAzol reagent?

Trypsinization is not needed. Simply add 1 mL of DNAzol Reagent per 10 cm2 of the culture plate area. Rock the plate back and forth and pipet the contents into a tube for ethanol precipitation of genomic DNA. If you want to trypsinize to count your cells, pellet the cells that will be used for DNA isolation after trypsinization. Remove the supernatant completely by aspiration, then add DNAzol Reagent to the cell pellet. Pipet 3 to 4 times to lyse the cells completely before proceeding to the centrifugation step.
The DNAzol procedure says to dissolve the genomic DNA in 8 mM NaOH. Will the 8 mM NaOH damage my DNA?

No, the 8 mM NaOH will not affect the DNA integrity. In fact, DNA is most stable at slightly alkaline pH (>7). You will find that the isolated DNA does not resuspend well in water and has even worse solubility in Tris buffer. (Water often has a pH of lower than 7 due to dissolved CO2 from the air. This slightly acidic water will actually cause degradation of your DNA!) The pH of the 8 mM NaOH is ~9 and can be easily adjusted with TE or HEPES once the DNA is in solution. (Over time, the solution becomes neutral upon exposure to air from dissolved carbon dioxide.)
Do I need to wash my cell pellet before DNAzol reagent treatment?

No. Washing is not necessary.
After DNAzol reagent extraction, my OD 260/280 ratio is low. Does this mean I have poor-quality DNA?

Consider the following if you have a low 260/280 ratio:

The correct amount of DNAzol reagent may not have been used. If DNAzol reagent was added to a cell pellet, make sure that the volume of reagent is 20 times that of the cell pellet.

There may have been a problem in pipetting away the viscous supernatant from the DNA pellet, leading to contamination with protein. The DNA may be used with DNAzol reagent again or extracted with phenol to remove the protein.

In some samples dissolved in water, the ratio may be low due to the acidity of the water or the low ion content in the water. The ratios may go up if the sample is dissolved in TE and the spec is zeroed with TE (or 1 to 3 mM Na2PO4, pH ~8.0). [See BioTechniques 22:474-6, 478-81 (1997).]. The molar extinction coefficient of the nucleotides is given at neutral pH, suggesting that the absorbance at 260 nm would be highest at neutral pH. Of course, DNA is not stable under acidic conditions so degradation may occur if the DNA is left in this condition for too long.

For Research Use Only. Not for use in diagnostic procedures.