Description
Silica membrane-based fast and convenient DNA purification from formalin-fixed paraffin-embedded (FFPE) tissue samples
The kit utilizes silica-based membrane technology in the form of a convenient spin column, eliminating the need for expensive resins, toxic phenol-chloroform extractions, or time-consuming alcohol precipitation. The standard procedure takes less than 20 minutes following cell lysis and yields purified DNA greater than 30kb in size. Isolated DNA can be used directly in PCR, Southern blotting and enzymatic reactions. Formalin-fixed and paraffin-embedded (FFPE) tissue samples are routinely prepared from human surgical tissue samples allowing to store them in archives for years. The Thermo Scientific GeneJET FFPE DNA Purification Kit is designed for fast and convenient purification of DNA from various amounts of archived FFPE tissue. Up to 8 sections (10 µm thickness) of FFPE sample from various tissues can be used for genomic DNA extraction without overnight incubation. Elimination of toxic reagents commonly used for deparaffinization (i.e., xylene) allows a safe DNA purification procedure with high DNA yields and excellent quality. Each preparation recovers up to 8 µg of genomic DNA that can be eluted in volumes from 20 µL allowing for concentrated samples. Due to high decrosslinking efficiency high quality purified DNA shows reliable performance in sensitive downstream applications such as qPCR, PCR (amplicons up to 1 kb) or NGS library preparation.
Highlights
•High quality DNA - suitable for PCR and qPCR
•Fast - procedure can be completed within 2 hours
•Safe - non toxic solvents
•Efficient - yields up to 8 µg of purified DNA
Applications
• PCR
• qPCR
• NGS library preparation
Includes
• Proteinase K Solution
• RNase A Solution
• Digestion Buffer
• Binding Buffer
• Wash Buffer I (conc.)
• Wash Buffer II (conc.)
• Elution Buffer
• GeneJET DNA Purification Columns & Collection Tubes
• Collection Tubes
• Detailed Protocol Hazardous:No
>Quality Control:
>•The GeneJET Genomic DNA Purification Kit is qualified by isolating genomic DNA from 200 µL of blood and 5 mg of mammalian tissue according to the recommended protocols.
>• The purified genomic DNA has an A260/280 ratio of ≥1.7.
>•A single band of more than 30 kb is seen after agarose gel electrophoresis and ethidium bromide staining.
>• Purified genomic DNA is evaluated by PCR amplification of a single-copy gene and digestion with FastDigest restriction enzymes.
> Storage Condition:Ambient Isolated genomic DNA is ideal for use in common molecular biology procedures, including:—PCR and qPCR, —FastDigest and conventional restriction digestion, —Southern blotting