MasterPure Complete DNA & RNA Purification Kit (Epicentre, Illumina Co)

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).

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 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.

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.

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.

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
Yeast(S. cerevisiae*)	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.

Ordering

Catalog No.		Concentration		Size

MasterPure™ Complete DNA and RNA Purification Kit

MC85200	200 Purifications*
MC89010	5/10 Purifications*

For isolating TNA, DNA, or RNA. Contents: Red Cell Lysis Solution, Tissue and Cell Lysis Solution, MPC Protein Precipitation Reagent, 2X T&C Lysis Solution, TE Buffer**, RNase A, RNase-Free DNase I, Proteinase K, 1X DNase Buffer, RiboGuard™ RNase Inhibitor. *Note: Cat. # MC89010 contains reagents for 10 TNA, 10 DNA, or 5 RNA purifications and Cat. # MC85200 contains reagents for 200 TNA, 200 DNA, or 100 RNA purifications.

MasterPure™ DNA Purification Kit (for isolating TNA or DNA)

MCD85201

200 Purifications

Contents: Red Cell Lysis Solution, Tissue and Cell Lysis Solution, MPC Protein Precipitation Reagent, 2X T&C Lysis Solution, TE Buffer**, RNase A, Proteinase K. **Not provided with the 10-purification kit (Cat. # MC89010).