About Quick-DNA™ Tissue/Insect Miniprep Kit
Note: The ZR Tissue & Insect DNA MiniPrep™ (D6016) has been renamed to Quick-DNA™ Tissue/Insect Miniprep Kit; all components and protocols are the same and have not been modified.The Quick-DNA™ Tissue/Insect Miniprep Kit is designed for the simple and rapid isolation of DNA (e.g., genomic, viral, mitochondrial) from fresh, frozen, or stored insect specimens including mosquitoes, bees, lice, ticks, and D. melanogaster. The procedure is easy and can be completed in minutes: samples are rapidly and efficiently lysed by bead beating with our state of the art, ultra-high density BashingBeads™. The DNA is then isolated and purified using our Fast-Spin column technology and is ideal for downstream molecular-based applications including PCR, array, genotyping, etc. The procedure is compatible with mammalian tissues, whole blood, and cultured cells.
|DNA Recovery||Typically, up to 25 μg total DNA is eluted into ≥ 25 μl DNA Elution Buffer per sample.|
|Processing Time||15 min|
|Equipment||Microcentrifuge, vortex, cell disruptor/pulverizer (optional).|
|DNA Size Limits||Capable of recovering genomic DNA up to and above 40 kb. In most instances, mitochondrial DNA and viral DNA (if present) will also be recovered.|
|Sample Sources||Samples (n ≥ 1 and ≤ 50 mg) of fresh, frozen, or stored insects including: mosquitoes, bees, lice, ticks, D. melanogaster, etc. Also, compatible with fresh or frozen mammalian tissues (e.g., soft tissues like liver and brain) as well as cultured cells, and whole blood.|
|DNA Purity||High quality DNA is eluted with DNA Elution Buffer that is well suited for PCR amplification, endonuclease digestion, etc. A260/A280 >1.8|
|DNA Yield||Expected yields can range from 1 - 5 µg DNA per mg specimen sampled. For mammalian tissues, yields are 1 - 3 µg DNA per mg of skeletal, heart and brain tissues and 3 - 5 µg DNA per mg of liver, kidney, and lung tissues. Whole blood will yield from 3 - 7 µg DNA per 100 µl.|
DNA yields from various insect and mouse samples using the Quick-DNA™ Tissue/Insect Miniprep Kit. Various amounts of sample were processed with equal volumes of eluted DNA analyzed in a 0.8% (w/v) agarose/ethidium bromide gel. The 1 kb DNA size marker is from Zymo Research
Add specimen(s) to a ZR BashingBead™ Lysis Tube. Add 750 µl Lysis Solution to the tube.
Generally, no more than 50 mg tissue should be sampled, for larger samples will exceed the DNA binding capacity of the spin column (See Specifications on page 1). Up to 400 µl of whole blood or up to 8.5 x106 cells suspended in 200 µl PBS can also be sampled.
Secure in a bead beater fitted with a 2 ml tube holder assembly (e.g., Disruptor Genie™) and process at maximum speed for 10 minutes.
Processing times may be as little as 40 seconds when using high-speed cell disrupters (e.g., the portable Xpedition™ Sample Processor, page 5, FastPrepâ-24, or similar). See manufacturer’s literature for operating information.
Centrifuge the ZR BashingBead™ Lysis Tube in a microcentrifuge at ≥10,000 x g for 1 minute.
Transfer up to 400 µl supernatant to a Zymo-Spin™ IV Spin Filter (orange top) in a Collection Tube and centrifuge at 7,000 rpm (~ 7,000 x g) for 1 minute.
Add 1,200 µl of Genomic Lysis Buffer to the filtrate in the Collection Tube from Step 4 and mix.
Transfer 800 µl of the mixture from Step 5 to a Zymo-Spin™ IIC Column in a Collection Tube and centrifuge at 10,000 x g for 1 minute.
Discard the flow through from the Collection Tube and repeat Step 6.
Add 200 µl DNA Pre-Wash Buffer to the Zymo-Spin™ IIC Column in a new Collection Tube and centrifuge at 10,000 x g for 1 minute.
Add 500 µl g-DNA Wash Buffer to the Zymo-Spin™ IIC Column and centrifuge at 10,000 x g for 1 minute.
Transfer the Zymo-Spin™ IIC Column to a clean 1.5 ml microcentrifuge tube and add 50 µl (25 µl minimum) DNA Elution Buffer directly to the column matrix. Centrifuge at 10,000 x g for 30 seconds to elute the DNA.