The MasterPure™ Complete DNA and RNA Purification Kit, MasterPure™ DNA Purification Kit, and MasterPure™ RNA Purification Kit use a novel, patent-pending technology that enables efficient purification of intact, high molecular weight DNA or RNA from a wide range of biological materials. Total nucleic acid (TNA), DNA, or RNA can be purified from bacteria, tissue culture cells, frozen or fresh mammalian tissue, paraffin-embedded tissue sections, mouse tail snips, and other biological materials. The protocol for TNA purification involves (1) an enzyme/salt/detergent-mediated cell lysis, (2) protein removal by precipitation, and (3) nucleic acid precipitation and resuspension. To isolate purified DNA, the sample is treated with RNase A; to isolate purified RNA, the sample is treated with RNase-Free DNase I. All manipulations are performed in standard microcentrifuge tubes. The recovered nucleic acids have an A260/280 ratio of 1.8-2.0, and can be used directly in PCR amplification (DNA and TNA) or RT-PCR (RNA). The standard MasterPure™ Complete DNA and RNA Purification Kit contains sufficient reagents for 200 DNA purifications or 100 DNA and 100 RNA purifications.
Targets | Samples Extracted |
Human & Mammalian |
Serum |
RNA & Genomic DNA |
Plasma |
HIV |
Whole blood |
E. coli |
Guthrie cards |
HCV |
Buccal cells |
B. pertussis |
Liver |
RSV |
Mouse tail |
Yeast |
Kidney |
M. tuberculosis |
Saliva |
Enterovirus |
Urine |
HPV |
Sputum |
Soy (tofu) |
Tissue culture cell lines |
Maize |
Cervical cells |
Insect tissues |
Paraffin-embedded tissues |
Streptococcus mutans |
|
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.
Figure 1. DNA, RNA, and total nucleic acid (TNA) purified from diverse cell sources using the MasterPure™ Complete Kit. M = kb ladder.
Figure 2. Basic MasterPure™ Complete Kit extraction protocol.
Sample | Sample Size | TNA µg | DNAµg | RNA µg |
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 |
0.1-0.2 |
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 |
2.2 x 106 cells |
|
|
11-18 |
(S. cerevisiae) |
1.1 x 107 cells |
|
|
>70-78 |
Table 2. Example of yields of nucleic acids obtained using MasterPure™ Complete DNA and RNA Purification Kits.
*For extraction of yeast DNA, the MasterPure Yeast DNA Purification Kit is recommended.
Figure 3. Consistent purification of DNA, total nucleic acid, and RNA. Five different genomic DNA (Lanes 1-5), total nucleic acid (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. M = kb ladder.
Figure 4. 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 the indicated number of E. coli cells.
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