The MasterPure™ Complete DNA and RNA Purification Kit enables rapid, high yield purification of high-molecular-weight genomic DNA, total cellular RNA and Total Nucleic Acid (TNA) from many different sample types.
The kit procedure uses a safe, gentle, and scalable salt-precipitation protocol (Figure. 1) that eliminates the need for hazardous chemicals and yield-reducing columns. Purification reactions can be scaled up or scaled down depending on the amount of nucleic acid needed and the size of the sample. MasterPure Kits for nucleic acid purification from Gram-positive bacteria, yeast (DNA or RNA), and blood samples are also available.
Applications
Purification of genomic DNA or total RNA for many applications, including:
- Library preparation for next generation sequencing (NGS) of genomic DNA and RNA
- DNA methylation studies using Illumina® Infinium® HumanMethylation BeadChips
- Genomic DNA and cDNA cloning
- qPCR and qRT-PCR
- Micrarray analyses (CGH, gene expression profiling, etc)
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Figure 1. Overview of the MasterPure Complete Kit protocol.
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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 × 10 7 (lanes 1) to 200 (lanes 6). Lane M, DNA marker; lane 7, negative control.
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Figure 3. DNA, RNA, and total nucleic acid (TNA) purified from diverse cell sources using the MasterPure Complete Kit. M = kb ladder.
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Targets
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Samples Extracted
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Human & mammalian RNA & genomic DNA HIV HCV RSV HPV Yeast Enterovirus
B. pertussis
E. coli
M. tuberculosis
Streptococcus mutans Soy (tofu) Maize Insect tissues
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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 analysed 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.
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Sample
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Sample Size
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TNA µg
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DNA µg
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RNA µg
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| HeLa/HL60 cells |
1 × 10 6 cells |
10-30 |
3-12 |
7-15 |
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Tissues
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| 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 |
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| Buffy coat |
300 µl |
40-55 |
40-55 |
3-6 |
| Mouse tail |
0.5 cm |
25-30 |
9-11 |
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E. coli
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3.5 × 10 6 cells |
2.5-2.8 |
1.3-1.6 |
1.6-1.8 |
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S. mutans
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1.5 ml |
0.9 |
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Yeast*
( S. cerevisiae)
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2.2 × 10 6 cells |
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11-18 |
| 1.1 × 10 7 cells |
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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.