About Genomic DNA Clean & Concentrator™
The Genomic DNA Clean & Concentrator™ (DCC™) is for the quick (5 minute) recovery of ultra-pure, large-sized DNA (e.g., genomic, mitochondrial, plasmid (BAC/PAC), viral, phage, (wga) DNA, etc.) from any enzymatic reaction or impure preparation (e.g., Proteinase K digestion). There is no need for organic denaturants, chloroform, or messy precipitations: simply add the specially formulated ChIP DNA Binding Buffer to a sample and then transfer the mixture to the supplied Zymo-Spin™ Column. Eluted DNA is suitable for sequencing, PCR, endonuclease digestion, and other enzymatic procedures. The product is also compatible with smaller DNAs (50 bp to 10 kb) from PCR, digestions, crude plasmid preparations, cDNA synthesis, etc.
|Format||Spin Column (capped)|
|DNA Recovery||Typically, up to 10 μg total DNA per column can be eluted with ≥10 μl water. Recovery of DNA ranges from 70 to 95%.|
|DNA Size Limits||Capable of purifying small DNA fragments >50 bp and large sized DNAs >200 kb.|
|Sample Sources||DNA from impure preparations of genomic DNA (e.g., Proteinase K digestions), plasmid DNA (including BAC), viral DNA, and whole genome amplified (wga) DNA. Can also be used for the purification of lower molecular weight DNA (50 bp to 10 kb) from PCR, endonuclease digestion, post-RT cDNA synthesis, etc.|
|DNA Purity||Ultra-pure (A(260/280) ≥ 1.8) high molecular weight DNA is eluted in water and is especially well suited for PCR, sequencing, endonuclease digestion, etc.|
|Product Detergent Tolerance||≤5% Triton X-100, ≤5% Tween-20, ≤5% Sarkosyl, ≤1% SDS|
Phage DNA Recovery. DNA (48.5 kb) is effectively recovered from 10-fold concentrations of starting material using the Genomic DCC™.
High molecular weight DNA is efficiently purified using the Genomic DCC™. Porcine gDNA (~35-50 kb), T4 phage DNA (170 kb), and DNA (48.5 kb) were purified (in duplicate) from input material using the Genomic DCC™. Eluted DNAs were analyzed in a 0.8% (w/v) TAE/agarose/EtBr gel (shown above). The size marker "M" is a 1 kb ladder (Zymo Research).
In a 1.5 ml microcentrifuge tube, add 2-5 volumes of ChIP DNA Binding Buffer to each volume of DNA sample1 (see table below). Mix thoroughly.
Transfer mixture to a provided Zymo-Spin™ IC-XL Column2 in a Collection Tube.
Centrifuge for 30 seconds. Discard the flow-through.
Add 200 µl DNA Wash Buffer to the column. Centrifuge for 1 minute. Repeat the wash step.
Add ≥ 10 µl DNA Elution Buffer3 or water4 directly to the column matrix and incubate at room temperature for one minute. Transfer the column to a 1.5 ml microcentrifuge tube and centrifuge at for 30 seconds to elute the DNA.
Ultra-pure DNA is now ready for use.
The Genomic DNA Clean & Concentrator™ was implemented for the purification of whole genome amplified DNA prior to generating DNA standards that are differentially methylated. This allowed accurate quantitation of methylation levels at the estrogen receptor alpha (ESR1) gene locus in the bovine endometrium