MasterPure™ Complete DNA and RNA Purification Kit

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

  1. 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.

 

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 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.
Sample Sample Size TNA µg DNA µg 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  
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.