Indispensable, for Protocols with Organic Extractions

• Increase Nucleic Acid Recoveries by up to 30%
• Eliminate Contamination with Interface Material
• Single Tube Enzymatic Reactions and Extractions
• Convenient Packaging for Extraction of Sample Volumes from 50 µl - 6 ml
• Reduce Exposure to Hazardous Organic Solvents

PHASE-Guard™, an inert proprietary gel, present during organic extractions migrates under centrifugal force to form a seal between the organic and aqueous phases. The organic phase and the interface material are effectively trapped below the PHASE-Guard™ seal. This barrier allows complete, easy transfer of the entire aqueous phase, by simply decanting or pipetting to a fresh tube.

Adding PHASE-Guard™ to your protocol allows for quantitative collection of the entire aqueous phase, resulting in a 20-30% increase in nucleic acid recovery. PHASE-Guard™ virtually eliminates contamination of the sample with interface material thus improving the quality of the purified nucleic acid. PHASE-Guard™ also offers the researcher increased protection from exposure to hazardous compounds.

Protocols Available:

• Genomic DNA
• Gel Extraction
• Mouse Tail DNA
• Total RNA
• Plasmid DNA
• Phage DNA

PHASE-Guard™ comes in two density formulations, Heavy and Light and can be adapted to virtually any protocol requiring extraction with phenol and/or chloroform by selecting the formulation that is compatible with the organic phase used (Table 1).

PHASE-Guard™ is inert, heat stable, and does not interfere with standard nucleic acid restriction and modification enzyme reactions. In fact, many of these reactions can be carried out directly in the 1.5 ml tubes containing PHASE-Guard™, terminated by the addition of phenol or phenol-chloroform and then extracted all in the same tube. PHASE-Guard™ can also be present in enzymatic reactions requiring a heat inactivation step (65°C for 10 minutes). Listed in Table 2 are restriction and modifying enzyme reactions that have been performed successfully in the presence of PHASE-Guard™.

PHASE-Guard™ is provided pre-dispensed for your convenience into various standard lab centrifuge tubes. Tube sizes available are 1.5 ml snap cap microcentrifuge tube; 15 ml screw cap conical tube, which can be centrifuged at up to 3,500 x g; and 12 ml snap cap round bottom tube, which can be centrifuged at up to 12,000 x g. These formats are for use with sample sizes ranging from 50 µl to 6 ml. PHASE-Guard™ also comes in 3 ml syringes for dispensing into a tube of your choice for any sample size.

By adding your sample to the PHASE-Guard™ tube, almost every standard laboratory procedure for nucleic acid isolation by aqueous/organic extraction (genomic DNA, plasmid DNA, total RNA, mRNA, phage or viral nucleic acid, etc.) is enhanced. Simply choose the PHASE-Guard™ density and tube size that fits with your protocol.

Table 1: PHASE-Guard™ Application Compatibility Guide. Aqueous Phase ORGANIC PHASE PCI CI H20 or Buffer Saturated PC H20 or Buffer Saturated Phenol £ 0.5 M NaCl LIGHT, HEAVY LIGHT, HEAVY LIGHT, HEAVY LIGHT £ 1 mg/mL BSA LIGHT, HEAVY LIGHT, HEAVY LIGHT, HEAVY LIGHT Cleared Bacterial Lysates HEAVY HEAVY HEAVY Not Recommended Tissue Homogenates LIGHT, HEAVY LIGHT, HEAVY LIGHT, HEAVY LIGHT RNA Purification HEAVY HEAVY HEAVY Not Recommended PCI = Phenol:Chloroform:Isoamyl Alcohol (25:24:1) CI = Chloroform:Isoamyl Alcohol (24:1) PC = Phenol:Chloroform (1:1)

Phase-Guard™ Procedure

Table 2: Retriction Enzymes Acc I EcoR V Nde I Sau3A I Ase I Hinc II Pac I Sma I BamH I Hind III Pme I Spe I Bgl I Hinf I Pst I Sph I Bgl II Hpa I Pvu II Sst I Cla I Kpn I Rsa I Xba I EcoR I Nco I Sal I Xmn I   Modifying Enzymes Calf Intestinal Alkaline Phosphatase T4 DNA Polymerase E. coli DNA Polymerase Reverse Transcriptase T4 Polynucleotide Kinase E. coli RNase H Terminal Deoxynucleotidyl Transferase T4 DNA Ligase E. coli DNA Ligase