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)
Figure 1. 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 × 10 7 (lanes 1) to 200 (lanes 6). Lane M, DNA marker; lane 7, negative control.
Figure 3. DNA, RNA, and total nucleic acid (TNA) purified from diverse cell sources using the MasterPure Complete Kit. M = kb ladder.
Targets Samples Extracted
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 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.
Sample Sample Size TNA µg DNA µg RNA µg
HeLa/HL60 cells 1 × 10 6 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 × 10 6 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 × 10 6 cells 11-18
1.1 × 10 7 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.
Key features
- Purify Total Nucleic Acid (TNA), DNA or RNA in 30-60 minutes
- Safe: Does not use hazardous phenol, chloroform or guanidine
- High Purity: A260/A280 ratios consistently between 1.8 and 2.0
- High Yields: Improves yields by avoiding the use of columns which often reduce nucleic acid yields
- Versatile: Purify TNA, genomic DNA, total RNA, FFPE RNA, or both genomic DNA and total RNA from a sample
- Total RNA Recovery: Purify both large and small (e.g., miRNA) RNA for RNA-Seq or qRT-PCR
- Proven: Hundreds of citations for purification of DNA and RNA from dozens of sample types for use in many applications
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