About ZR Genomic DNA™ -Tissue MicroPrep
The ZR Genomic DNA™-Tissue MicroPrep 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.
|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.|
High yield/quality DNA is successfully isolated from porcine muscle using the ZR Genomic DNA™-Tissue MiniPrep. Equivalent amounts (25 mg) of muscle tissue were processed using the ZR Genomic DNA™ Tissue MiniPrep after incubation with Proteinase K at 55ºC for the indicated times (in minutes) or overnight (O/N). Equal volumes of eluted DNA were analyzed in a 0.8% (w/v) TAE/agarose/ethidium bromide gel. M: 1 kb ladder (Zymo Research).
To a tissue sample (≤5 mg) in a microcentrifuge tube and 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™ IC 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 ≥10 µl DNA Elution Buffer or water (e.g., add 40 µl if sampling 5 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.
Genomic DNA was isolated from the Caribbean dipsadid snake using the ZR Genomic DNA™ - Tissue MicroPrep; the isolated DNA was then used for PCR and mitochondrial and nuclear loci sequencing. Phylogenetic analysis confirms the morphology and its identification as the Cuban Racer, a species previously regarded as endemic to Cuba.
The ZR Genomic DNA™ - Tissue MicroPrep was used to isolate DNA from different anole specimens followed by PCR and sequencing. Phylogenetic analysis suggested that all samples have mtDNA originating from a native species yet the possibility of a new nonnative introduced anole cannot be ruled out.