Takara's Dr. GenTLE Systems are designed to enable the extraction of highly purified DNA from whole blood (9081) for use in various reactions. The standard protocol describes extraction from 100 µL of whole blood, but the kit can also be used for larger-volume extraction. The protocol is suitable for simultaneously processing a large number of samples.
The Dr. GenTLE whole blood kits are composed of three solutions. GenTLE Solution I lyses cells immediately after being added to the sample, and a noncharged complex is formed with nucleic acids. The complex is then collected by centrifugation, and the precipitate washed with GenTLE Solution II. Dr. GenTLE Solution III is added to isolate only DNA, which can be precipitated by the addition of isopropanol. The protocol takes just 40 minutes and yields highly purified DNA.
Dr. GenTLE High Recovery (from Yeast) (9082) is designed to efficiently extract and purify genomic DNA from yeast cells by using both a digestive enzyme of the cell wall and salting out for DNA purification. This kit is useful to isolate genomic DNA, which is available for general genetic engineering, such as Southern blotting, PCR, or restriction enzyme treatment.
Using the whole blood kit for blood samples: Generally, 2–7 µg of DNA will be obtained from 100 µL of sample, irrespective of the anticoagulant used. The purity of the obtained DNA is A260/280 = 1.7–2.0.
Using the high recovery yeast kit for yeast: For standard experiment, 4–10 µg of genomic DNA (35–200 kb) can be obtained from 1–2 x 108 yeast cells.
Use of Dr. GenTLE Precipitation Carrier in ethanol precipitation results in higher recoveries of dilute (5 ng/mL) DNA samples than glycogen. Also, low temperature incubation is not required, and the white pellet generated is easily visible, preventing loss during washing.
