About Quick-DNA™ Tissue/Insect 96 Kit
Note: The ZR-96 Tissue & Insect DNA Kit™ (D6017) has been renamed to Quick-DNA™ Tissue/Insect 96 Kit; all components and protocols are the same and have not been modified.The Quick-DNA™ Tissue/Insect 96 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 plate 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 5 µg total DNA is eluted into ≥ 25 µl DNA Elution Buffer per sample.|
|Processing Time||40 min|
|Equipment||Centrifuge with microplate carriers, 96-well plate/block disruptor or pulverizer.|
|DNA Size Limits||> 1 kb|
|Sample Sources||Samples (n ≥ 1 and ≤ 10 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 DN|
DNA yields from various insect and mouse samples using the Quick-DNA™ Tissue/Insect 96 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 the tubes of a ZR BashingBead™ Lysis Rack. Add 400 µl Lysis Solution to the tube. Cap tubes tightly to prevent leakage.
Generally, no more than 10 mg tissue should be sampled, for larger samples will exceed the DNA binding capacity of the Silicon-A™ Plate (See Specifications on page 1). Up to 100 µl of whole blood or up to 1.7 x106 cells suspended in 100 µl PBS can also be sampled.
Secure in a 96-well block/plate bead beater (e.g., 2010 GenoGrinder®) and process samples. Optimization of processing time/speed will be necessary for complete sample lysis.
Processing times may be as little as one minute when using high-speed bead beaters (e.g., 2000 GenoGrinderâ, page 4). See manufacturer’s literature for specific operating information.
Centrifuge the ZR BashingBead™ Lysis Rack at ≥ 3,000 x g (5,000 x g max.) for 5 minutes.
Transfer up to 250 µl supernatant to each well of a 96-Well Block.
Add 750 µl of Genomic Lysis Buffer to the supernatant in the 96-Well Block from Step 4. Cover completely with Cover Foil and mix thoroughly by vortexing block for 2 minutes. Centrifuge the 96-Well Block at ≥ 3,000 x g (5,000 x g max.) for 5 minutes.
Remove or pierce foil and transfer 500 µl of each of the supernatants from Step 5 to the wells of a Silicon-A™ Plate on a Collection Plate. Centrifuge the assembly at ≥ 3,000 x g (5,000 x g max.) for 5 minutes.
Discard the flow through from the Collection Plate and repeat Step 6.
Add 200 µl DNA Pre-Wash Buffer to the wells of the Silicon-A™ Plate on the emptied Collection Plate and centrifuge the assembly at ≥ 3,000 x g for 5 minutes.
Add 500 µl g-DNA Wash Buffer to the wells of the Silicon-A™ Plate on the Collection Plate and centrifuge the assembly at ≥ 3,000 x g for 5 minutes.
Transfer the Silicon-A™ Plate to an Elution Plate and add 100 µl (25 µl minimum) DNA Elution Buffer directly to the matrices in the plate. Centrifuge the assembly at ≥ 3,000 x g for 5 minutes.
Eluted, ultra-pure DNA is now ready for use in your experiments, or the Elution Plate can be covered with Cover Foil for storage of the DNA.