About ZR Tissue & Insect DNA MidiPrep™
Note: This product will be discontinued within the next 3 months (November 30th, 2016) or while supplies last. Please refer to ZR Tissue & Insect DNA MiniPrep (D6016) for the replacement product.The ZR Tissue & Insect DNA MidiPrep™ 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 ~125 µg total DNA is eluted into ≥ 150 µl DNA Elution Buffer per sample.|
|Processing Time||20 min|
|Equipment||Centrifuge, Vacuum Source and Manifold, Microcentrifuge, Cell Disrupter/Pulverizer w/ 50 ml Tube Adapter|
|DNA Size Limits||Capable of recovering genomic DNA sized fragments from 100 bp to ≥40 kb. Typical fragment sizes range from 25 to 35 kb. If present, parasitic and viral DNA will also be recovered.|
|Sample Sources||Samples (n ≥2.5 and ≤125 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, humic/fulvic-free DNA is eluted with DNA Elution Buffer making it perfect for PCR. A260/280 >1.8, A260/230 >2.0|
|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 ZR Insect & Tissue DNA MiniPrep™. 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 specimens(s) to a ZR BashingBead™ Lysis/Filtration Tube1. Add 6 ml Genomic Lysis Buffer to the sample, cap tube1, and process.
(To prevent the Lysis Solution from leaking into the bottom of the 50 ml tube, place the ZR BashingBead™ Lysis/Filtration Tube on its side prior to processing).
Generally, no more than 125 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 2 ml of whole blood or up to 3x107 cells suspended in 1 ml PBS can also be sampled.
Secure in a bead beater fitted with a 50 ml tube holder assembly (see page 5) to process samples. Optimization of processing time/speed will be necessary for complete sample lysis.
Processing times may be as little as 40 seconds when using high-speed cell disrupters (e.g., FastPrepâ-24, Geno/Grinderâ, page 5). See manufacturer’s literature for operating information.
Centrifuge the ZR BashingBead™ Lysis/Filtration Tube in a centrifuge at ≥3,000 x g (5,000 x g max.) for 5 minutes.
Filter the entire mixture from Step 3 using a Zymo-Spin™ V-E Column/Zymo-Midi Filter™ assembly mounted on a vacuum manifold2,3 (see diagram on page 2) with a vacuum source set at ≥600 mm Hg.
Disconnect the Zymo-Spin™ V-E Column/Zymo-Midi Filter™ assembly and transfer the Zymo-Spin™ V-E Column to a Collection Tube. Spin at 10,000 x g for 1 minute in a microcentrifuge4, then add 300 µl DNA Pre-Wash Buffer to the column and spin at 10,000 x g for 1 minute. Discard the flow through.
Add 400 µl g-DNA Wash Buffer to the column and centrifuge at 10,000 x g for 1 minute. Discard flow through and repeat wash step.
Transfer the Zymo-Spin™ V-E Column to a 1.5 ml microcentrifuge tube and add 150 µl DNA Elution Buffer directly to the column matrix5 and allow column to stand for 1 minute at room temperature. Centrifuge at 10,000 x g for 1 minute to elute the DNA.
Ultra-pure DNA is now ready for use in your experiments.