5 PRIME Phase Lock Gel:
Convenient and efficient phenol or phenol/chloroform extractions of nucleic acids, plasmid, genomic DNA and RNA.
5 PRIME Phase Lock Gel (PLG) is available in Australia through Gene Target Solutions. It is a unique product that eliminates interphase-protein contamination during phenol extraction and ensures faster results with improved recoveries. Phase Lock Gel 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.
- For organic extraction methods
- Forms stable barrier between phases
- Pre-aliquoted in light and heavy variants
- PLG Heavy compatible with Isol-RNA Lysis reagent
- Gel barrier allows easy sample decanting
- Reduced contact with hazardous organic solvents
- Improved performance of homebrew
- Eliminates interphase contamination of nucleic acid solution
- 30% greater yield of nucleic acids over conventional method
- Compatible with most protocols
- Best choice for fat-rich samples
5 Prime Phase Lock Gel can be adapted to virtually any protocol requiring extraction of an aqueous sample with phenol and/or chloroform. For convenience, Phase Lock Gel is provided predispensed into standard 2mL centrifuge tubes and comes in two varieties:
Phase Lock Gel 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).
Phase Lock Gel Light
Aqueous phase: Low-density sample
Organic phase: Standard mixtures of phenol, chloroform, isoamyl alcohol.
Phase Lock Gel 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 Phase Lock Gel at the appropriate temperature and then terminated and extracted by the addition of phenol or phenol/chloroform to the reaction tube. Phase Lock Gel 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.
Phenol Extraction – The Challenges
- Highly volatile
- Highy toxic
- Recovery of aqueous phse
- Need for cautious pipetting
- Less standardisation
- Yield loss due to incomplete recovery of aqueous phase
- Carryover of organic phase
- Risk of inhibition of downstream application
- Less standardisation
Contents: 200 2mL tubes
- 2302820 – Phase Lock Gel Light 2ml
- 2302830 – Phase Lock Gel Heavy 2ml
Storage & Handling:
Store at room temperature; do not refrigerate or freeze.