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View additional product information for DNAzol™ Reagent, for isolation of genomic DNA from solid and liquid samples - FAQs (10503027)
16 product FAQs found
如果DNAzol试剂的用量过少，则可能在加入乙醇后出现两相。应加入更多DNAzol试剂并继续实验。
如果在分离DNA前使用磷酸盐缓冲液洗涤细胞或组织，则磷酸盐可能会抑制限制性内切酶。我们推荐向DNA溶液中加入DNAzol试剂，使用0.5倍体积的95%乙醇再次沉淀。沉淀后，使用95%乙醇洗涤2次，短暂干燥，并重悬于8 mM NaOH中。
您可以尝试将样本溶于8 mM NaOH中，在37°C孵育过夜以帮助DNA的溶解。您也可以尝试在45°C孵育15分钟。
可以，这种情况是由试剂中的染料所致，并且可能取决于储存试剂的体积。颜色改变不会影响性能。
使用DNAzol试剂分离得到的实际上是总DNA，所以质粒DNA也会和基因组DNA一并分离出来。线粒体基因组与质粒类似，也可通过DNAzol试剂分离出来。可将离心步骤之前的乙醇室温孵育步骤从1分钟延长至5-10分钟，以获得最大的DNA得率。
我们推荐将DNAzol试剂储存于室温。DNAzol的裂解物（匀浆）可于15–30°C下储存1个月；在4°C或–20°C条件下储存10个月之后，DNAzol裂解物（匀浆）中仍能够提取得到高分子量的、可被限制性内切酶完全酶切的基因组DNA，且DNA在PCR反应中也可以得到较好的结果。在漂洗阶段，DNA可以保存于95%的乙醇中，在15°C至30°C至少储存一周，或4°C左右条件下至少储存三个月。DNA保存于DNAzol试剂中，可于室温下储存一个月，或4°C下储存十个月。
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.
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.
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.
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.
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.
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.
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.
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.)
No. Washing is not necessary.
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.