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Nucleic Acid Purification
GeneJET™ Genomic DNA Purification Kit
- Product information
-
Overview of purification procedure.
- Features
- Universal – can be used for both cell and tissue samples from a wide range of sources.
- Efficient – high yields of high molecular weight genomic DNA.
- Fast – 20 minute procedure following cell lysis.
- Convenient – spin columns are capped and assembled with collection tubes.
- Applications
- Isolated genomic DNA is ideal for use in common molecular biology procedures, including:
– PCR and qPCR,
– FastDigest® and conventional restriction digestion,
– Southern blotting.
- Isolated genomic DNA is ideal for use in common molecular biology procedures, including:
DescriptionThe 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 (see Fig.1, Table 1).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 30 kb in size. Isolated DNA can be used directly in PCR, Southern blotting and enzymatic reactions.
Quality ControlThe 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.
Components- Proteinase K Solution
- RNase A Solution
- Digestion Solution
- Lysis Solution
- Wash Buffer I (concentrated)
- Wash Buffer II (concentrated)
- Elution Buffer
- GeneJET™ Purification Columns (pre-assembled with collection tubes)
- Collection tubes
- Detailed Protocol
Figure 1. Genomic DNA purified from various sources using the GeneJET™ Genomic DNA Purification Kit.
150 ng of isolated genomic DNA from each source was fractionated by agarose gel (0.5%) electrophoresis.
M – Lambda DNA/HindIII Marker (#SM0101)
1 – human bone marrow
2 – human blood
3 – rat spleen
4 – mouse heart
5 – mouse liver
6 – HeLa cells
7 – Jurkat cells
8 – E.coli DH5alfa cells
Figure 2. Yields of genomic DNA purified using the GeneJET™ Genomic DNA Purification Kit and kits from other vendors.
Table 1. Typical genomic DNA yields from various sources.Source Quantity Yield, µg Mammalian blood 200 µl 4-6 Mouse heart 10 mg 10-15 Mouse tail 0.5 cm 8-10 Rat liver 10 mg 10-20 Rat spleen 5 mg 20-30 Rat kidney 10 mg 25-30 Rabbit ear 20 mg 5-10 Bacillus pumilis cells 2x109 cells 10-15 Escherichia coli cells 2x109 cells 10-15 HeLa cells 2x106 cells 15-20 Jurkat cells 5x106 cells 25-30 Saccharomyces cerevisiae cells 1x108 cells 3-5
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