About ZR-96 Genomic DNA™ -Tissue MiniPrep
Note: Catalog numbers D3055, D3056, and D3057 will be discontinued within the next 3 months (March 31st, 2017) or while supplies last. Please refer to the D4070 and D4071 for the replacement products.
The ZR-96 Genomic DNA™-Tissue MiniPrep provides a simple, high-throughput procedure for the rapid isolation of total DNA (e.g., genomic, mitochondrial, parasitic, microbial, viral) from a variety of solid tissues. The product has been optimized for maximal recovery of ultra-pure DNA without RNA contamination and is also compatible with inputs including buffy coat, bone marrow, cells from culture, whole blood (fresh or stored), serum, plasma, and many biological liquid samples. For processing, simply digest the sample with the supplied Proteinase K then add the Genomic Lysis Buffer, vortex, and transfer the mixture to the supplied Silicon-A™ plate. PCR inhibitors are effectively removed during the purification process and purified DNA is suitable for downstream applications including: PCR, Southern blotting, sequencing, endonuclease digestion, bisulfite conversion/methylation analysis, etc.
|Processing Time||45 min|
|Equipment||Water bath or heat block (55°C), Microcentrifuge, and vortex.|
|DNA Size Limits||Capable of recovering genomic and mitochondrial DNA sized fragments from 100 bp to ≥ 40 kb. If present, Parasitic, microbial, and viral DNA will also be recovered. Typical fragment sizes range from 25 kb - 35 kb.|
|Sample Sources||Solid tissues (e.g., tailsnips, earpunches, adipose tissue, etc.), whole blood, plasma, serum, buffy coat, lymphocytes, cultured cells, buccal cells, FFPE tissues, semen, hair, and other biological sources are effectively processed using this kit.|
|DNA Purity||High quality DNA for PCR, endonuclease digestion, Southern blotting, bisulfite conversion/methylation detection, sequencing, genotyping, etc., is eluted with DNA Elution Buffer or water (A260/A280 ≥1.8).|
|Product Detergent Tolerance||≤5% Triton X-100, ≤5% Tween-20, ≤5% Sarkosyl, ≤0.1% SDS|
|DNA Yield||The DNA binding capacity of the plate is 5 μg/well. Typically, mammalian tissues yield: 1 - 3 µg DNA per mg skeletal, heart, and brain tissues and 3 - 5 µg DNA per mg liver, kidney and lung tissues. Human whole blood will yield 3 - 7 µg DNA per 100 µ|
To each tissue sample (≤ 10 mg) in a well of a Deep-Well Block add a solution of...
H2O 70 µl
2X Digestion Buffer 70 µl
Proteinase K 10 µl
Mix, seal blocks with film, and then incubate at 55oC for 1-3 hours.
Note: If required (FFPE samples), digesting samples overnight at 55oC with Proteinase K is possible without affecting the integrity of the DNA
Add 450 µl Genomic Lysis Buffer to each of the wells and mix thoroughly by repeated pipetting. Centrifuge blocks at ≥1,000 x g for 5 minutes to remove insoluble debris.
Transfer the supernatants to the wells of a Silicon-A™ Plate on a Collection Plate. Centrifuge at ≥2,500 x g (5,000 x g max.) for 5 minutes.
Add 200 µl of DNA Pre-Wash Buffer to each well and centrifuge at ≥2,500 x g for 5 minutes. Discard the flow through.
Add 300 µl of g-DNA Wash Buffer to each well and centrifuge at ≥2,500 x g for 5 minutes.
Transfer the Silicon-A™ Plate onto a Collection Plate. Add ≥30 µl DNA Elution Buffer or water to each well. Incubate 2-5 minutes at room temperature, then centrifuge at ≥2,500 x g for 5 minutes to elute the DNA. The eluted DNA can be used immediately for molecular based applications or stored ≤-20ºC for future use.