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

otal RNA separated on a denaturing gel will have sharp, clear 28S and 18S rRNA bands (eukaryotic samples). The 28S rRNA bands of the samples used with or without PLG are approximately twice as intense as the 18S rRNA band (Fig. 2). This 2:1 ratio (28S:18S) is a good indication that the RNA is completely intact.

Fig. 2:. Total RNA were separated by gel electrophoresis beside the Gene Ruler DNA Marker 1010,000 bp (Fermentas) on a 1% denaturing agarose gel.

Lane 1: Total RNA prepared with the use of Eppendorf Phase Lock Gel. Lane 2: Total RNA prepared without the use of Eppendorf Phase Lock Gel. Reverse Transcription PCR

In each preparation, an 844 bp fragment of the human p53 cDNA could be amplified from RNA prepared with or without the use of PLG. As shown in Fig. 3, the amplicons are of the same size and approximately the same intensity, and there are no additional bands visible.

Fig. 3: RT-PCR performed on total RNA isolated with TRIzol and Phase Lock Gel. '"/>

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