About ZR Genomic DNA™ -Tissue MiniPrep
Note: Catalog numbers D3050 and D3051 will be discontinued within the next 3 months (March 31st, 2017) or while supplies last. Please refer to D4068 and D4069 for the replacement products.
The ZR Genomic DNA™-Tissue MiniPrep is a simple 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 are 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 spin column. PCR inhibitors are effectively removed during the purification process and purified DNA is suitable for downstream applications including: PCR, Southern blotting, DNA sequencing, endonuclease digestion, bisulfite conversion/methylation analysis, etc.
|Processing Time||25 min|
|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 column is 25 µg. 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 µl blood sampled. DNA is eluted into ≥ 50 µl DNA Elution Buffer or water.|
|Equipment Needed||Water bath or heat block (55°C), microcentrifuge, and vortex.|
|DNA Size||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.|
To a tissue sample (≤ 25 mg) in a microcentrifuge tube add a solution of...
H2O 95 µl
2X Digestion Buffer 95 µl
Proteinase K 10 µl
Mix and then incubate the tube at 55oC for 1-3 hours.
Note: If required (e.g., FFPE samples), digesting samples overnight at 55oC with Proteinase K is possible without affecting the integrity of the DNA.
Add 700 µl Genomic Lysis Buffer to the tube and mix thoroughly by vortexing. Centrifuge at 10,000 x g for one minute to remove insoluble debris.
Transfer the supernatant to a Zymo-Spin™ IIC Column in a Collection Tube. Centrifuge at 10,000 x g for one minute.
Add 200 µl of DNA Pre-Wash Buffer to the spin column in a new Collection Tube. Centrifuge at 10,000 x g for one minute.
Add 400 µl of g-DNA Wash Buffer to the spin column. Centrifuge at 10,000 x g for one minute.
Transfer the spin column to a clean microcentrifuge tube. Add ≥50 µl DNA Elution Buffer or water (e.g., add 200 µl if sampling 25 mg tissue) to the spin column. Incubate 2-5 minutes at room temperature, then centrifuge at top speed for 30 seconds to elute the DNA. The eluted DNA can be used immediately for molecular based applications or stored ≤-20ºC for future use.