DNA extraction from Blood Through Kit Method
STORAGE
Proteinase K and RNase A solutions are stable at room temperature as long as not opened.
After being opened they should be stored at -20 °C. Other components of the kit should be
stored at room temperature (15-25 °C).
Note. Close the bag with GeneJET Genomic DNA Purification Columns tightly after each use!
DESCRIPTION
The GeneJET™ Genomic DNA Purification Kit is designed for rapid and efficient purification of
high quality genomic DNA from various mammalian cell culture and tissue samples, whole
blood, bacteria and yeast. 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 of more than 30 kb in size. Isolated DNA
can be used directly in PCR, Southern blotting and enzymatic reactions. See Table 1 for
typical genomic DNA yields from various sources.
PRINCIPLE
Depending on the starting material, samples are digested with Proteinase K in either the
supplied Digestion or Lysis Solution. RNA is removed by treating the samples with RNase A.
The lysate is then mixed with ethanol and loaded on the purification column where the DNA
binds to the silica membrane. Impurities are effectively removed by washing the column with
the prepared wash buffers. Genomic DNA is then eluted under low ionic strength conditions
with the Elution Buffer
Buffers
For mammalian cell lysate preparation:
PBS (137 mM NaCl, 2.7 mM KCl, 10 mM Na2HPO4, 2 mM KH2PO4, pH 7.4)
TE buffer (10 mM Tris-HCl, pH 8.0, 1 mM EDTA)
For gram-positive bacteria lysate preparation
Gram-positive bacteria lysis buffer (20 mM Tris-HCl, pH 8.0, 2 mM EDTA,
1.2% Triton X-100, add lysozyme to 20 mg/mL immediately before use)
For yeast lysate preparation:
Yeast lysis buffer (5 mg/mL zymolyase 20T, 1 M sorbitol, 0.1 M EDTA