Sample Preparation |
Purification of Genomic DNA
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Applications
- Purification of TNA, DNA, or RNA for many applications, including PCR,
RT-PCR, cloning, sequencing, Northern and Southern blotting, restriction
analysis, and genomic library preparation.
The MasterPure™ Complete DNA and RNA Purification Kit uses a novel technology
that enables efficient purification of intact total nucleic acid (TNA), DNA,
or RNA from every type of biological material (Table 1 and Fig. 1).
The gentle salt-precipitation protocol permits rapid purification of high-molecular-weight
nucleic acids from a single sample or many samples that are processed simultaneously
(Fig. 4). High yields of nucleic acids are obtained using MasterPure Kits (Table
2), and yields and purity are highly consistent for different samples of the
same material (Fig. 2). The resultant nucleic acids have an A260/A280 ratio
of 1.8-2.0, indicating that they are substantially free of protein. The TNA,
DNA, and/or RNA purified from different samples can be used directly in many
applications. For example, the sensitivity of PCR or RT-PCR assays is often
improved if the template is obtained using a MasterPure Purification Kit.
Benefits
- Purify TNA in 30 minutes without cumbersome columns.
- A260/A280 ratios consistently between 1.8 and
2.0.
- Recover >90% of theoretical DNA yield.
- No phenol, chloroform, or other caustic solvents.
- Each kit component is also available separately in a 10X package
size.
- Low- and high-molecular-weight nucleic acids are recovered.
Reference
- Schanke, J. and Watson, J. (1998) EPICENTRE Forum 5(2),
12.
|
Figure 1 (click to enlarge). Overview of the MasterPure™ Complete
Kit protocol. |
Figure 2. Comparison of PCR sensitivity using templates obtained
using the MasterPure™ Complete DNA and RNA Purification Kit versus
a spin-column kit. PCR amplification was performed after extraction
from serial dilutions of E. coli cells ranging from 2 x 107 (lanes
1) to 200 (lanes 6). Lane M, DNA marker; lane 7, negative control. |
Figure 3 (click to enlarge). DNA, RNA, and total nucleic acid
(TNA) purified from diverse cell sources using the MasterPure™ Complete
Kit. M = kb ladder. |
Human & mammalian RNA & genomic DNA
HIV
HCV
RSV
HPV
Yeast
Enterovirus
B. pertussis
E. coli
M. tuberculosis
Streptococcus mutans
Soy (tofu)
Maize
Insect tissues |
Plasma
Whole blood
Guthrie cards
Buccal cells
Liver
Mouse tail
Kidney
Saliva
Urine
Sputum
Tissue-culture cell lines
Cervical cells
Paraffin-embedded tissues |
Table 1. Examples of targets and samples analyzed using MasterPure™ DNA
and RNA Purification Kits. Using an extremely simple salt-precipitation
protocol, TNA, DNA, or RNA can be isolated from virtually any source
in less than 1 hour. |
| HeLa/HL60 cells |
1 X 106 cells |
10-30 |
3-12 |
7-15 |
| Tissues |
| Liver |
5 mg |
33-42 |
5-10 |
13-25 |
| Brain |
5 mg |
9-13 |
6-9 |
4-11 |
| Heart |
5 mg |
6-10 |
4-7 |
4-5 |
| Kidney |
5 mg |
10-17 |
3-8 |
14-17 |
| Thymus |
5 mg |
15-30 |
6-12 |
9-18 |
| Blood |
200 µl |
3-10 |
3-9 |
|
| Buffy coat |
300 µl |
40-55 |
40-55 |
3-6 |
| Mouse tail |
0.5 cm |
25-30 |
9-11 |
|
| E. coli |
3.5 x 106 cells |
2.5-2.8 |
1.3-1.6 |
1.6-1.8 |
| S. mutans |
1.5 ml |
0.9 |
|
|
Yeast*
(S. cerevisiae) |
2.2 x 106 cells |
|
|
11-18 |
| 1.1 x 107 cells |
|
|
70-78 |
*For extraction of yeast DNA, the MasterPure™ Yeast DNA Purification
Kit is recommended.
Table 2. Example of yields of nucleic acids obtained using MasterPure™ Complete
DNA and RNA Purification Kits. |
Figure 4. Consistent purification of DNA, TNA, and RNA. Five
different genomic DNA (lanes 1-5), TNA (lanes 6-10), and total cellular
RNA (lanes 11-15) samples were isolated from frozen bovine liver with
the MasterPure™ Complete DNA and RNA Purification Kit. Lane M,
Kilobase ladder. |
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MasterPure™ RNA Purification Kit
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