5PRIME Phase Lock Gel (PLG) is a unique product that eliminates interphase-protein contamination during phenol extraction and ensures faster results with improved recoveries. PLG migrates to form a tight seal between the phases of an aqueous/organic extraction during centrifugation. The organic phase and the interphase materials are effectively trapped in or below the barrier, thus enabling complete and easy decanting or pipetting of the entire aqueous phase. The benefits are increased yields of up to 30%, increased protection from exposure to hazardous compounds, and no risk of interphase sample contamination.
PLG can be adapted to virtually any protocol requiring extraction of an aqueous sample with phenol and/or chloroform1. For convenience, PLG is provided predispensed into standard 2mL centrifuge tubes and comes in two varieties:
PLG Heavy
Aqueous phase: High density sample (for example, high salt content).
Organic phase: Standard mixtures of phenol, chloroform, isoamyl alcohol with a chloroform fraction of at least 60% (Phenol as the sole organic solvent is not compatible with PLG Heavy).
PLG Light
Aqueous phase: Low-density sample
Organic phase: Standard mixtures of phenol, chloroform, isoamyl alcohol.
PLG is inert and stable to mild heating, and it does not interfere with any of the standard nucleic acid restriction and modification enzymes. The reactions can be carried out in the presence of PLG at the appropriate temperature and then terminated and extracted by the addition of phenol or phenol/chloroform to the reaction tube. PLG can be present during the heating stage of the deactivation of enzymes (65°C for 10 min.) prior to the organic extraction.
Genomic DNA from Mouse Tails: Use PLG Heavy to enhance isolation of genomic DNA from mouse tails using a standard proteinase K/SDS organic extraction protocol.
Plasmid DNA Isolation: Use PLG Heavy to enhance isolation of plasmid DNA using standard alkaline lysis/phenol extraction protocols.
Total RNA Isolation: Use PLG Heavy to enhance isolation of total RNA from cells and tissues using a modified guanidinium isothiocyanate/acid phenol method.
Genomic DNA Isolation: Use PLG Light to enhance isolation of genomic DNA using standard proteinase K/SDS procedures.
DNA Isolation from Agarose Gels: Use PLG Light to enhance isolation of DNA fragments from agarose gels using phenol extraction procedures.
Lamba Phage or M13 DNA Isolation: Use PLG Light to enhance isolation of bacteriophage DNA using standard purification protocols.
200 2mL tubes
5PRIME Phase Lock Gel is stable when stored at room temperature (DO NOT FREEZE). Phase Lock Gel is inert and does not interfere with standard nucleic acid restriction and modification enzymes. Phase Lock Gel can be present during the heat inactivation of enzymes (65°C for 10 minutes) prior to organic extraction. For lot specific expiry date, refer to package label, Certificate of Analysis or Product Specification Form.
User Manual