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Easy preparation of total RNA from cultured cells with TRIzol and ,,, Eppendorf Phase Lock Gel

body> Lanes 15: 844 bp fragment of the human p53 gene amplified by RT-PCR from total RNA prepared with TRIzol and with Eppendorf Phase Lock Gel Lanes 610: 844 bp fragment of the human p53 gene amplified by RT-PCR from total RNA prepared with TRIzol but without Eppendorf Phase Lock Gel Lane 11: Negative control, RT-PCR murine RNA Lane 12: 0.5 mg of DNA size marker (Invitrogen) Range: 506 to 21,216 bp Conclusion

Where it is used as a standard helpful tool for the preparation of DNA via phenol extraction, the Eppendorf Phase Lock Gel (PLG) Heavy can also be used in combination with mono-phasic extraction reagents such as TRIzol for the preparation of total RNA from tissue culture cells. The use of Eppendorf PLG increases the yield of isolated RNA and shortens the preparation time. With the help of PLG the separation of the aqueous and the organic phase is more convenient and reliable and the risk of losing the sample due to organic impurities is minimized (i.e. recovery of the entire aqueous phase is possible).

The RNA is of high purity and not contaminated with proteins or organic solvents measured by UV-absorption. Total RNA prepared with Eppendorf Phase Lock Gel can be directly used for downstream applications sensitive to any kind of impurities in the preparation. Reverse transcription of RNA into cDNA and subsequent amplification could be performed successfully.

In summary, Eppendorf Phase Lock Gel (PLG) Heavy is an excellent tool for simplifying the pre
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