For semi-quantitative RT-PCR (sqRT-PCR), Trizol isolated RNA was cleaned with the RNeasy Plant Mini Kit (Qiagen) and a DNase I digest following the manufacturers instructions was performed.
After RNA isolation, the samples were treated with recombinant RNase-free DNase I with Protector RNase inhibitor (Roche Applied Science, Penzberg, Upper Bavaria, Germany) according to the manufacturer's instructions to digest any remaining DNA.
An aliquot was treated with Terminator enzyme (Epicentre® Biotechnologies) according to the manufacturer's instructions to selectively digest 5'-monophosphate transcripts.
DNA samples were extracted from the cells by the Genomic DNA purification kit (Gentra, Minneapolis, MN, USA) according to the manufacturer's instructions, digested for 16 h by HinfIII and RSAI and separated on 0·8% agarose gels.
"Without a lot of instruction, he was able to digest a lot of information.
The educators believe a full day of kindergarten allows more in-depth instruction and gives students time to digest what they are learning.
In my experience, most students are quite able to read and digest a practical schedule and follow suitably clear instructions.
Extraction of total RNA was performed using the Qiagen RNeasy kit (Qiagen, Hilden, Germany) including a DNA digest on-column according to the manufacturer's instructions.
Total RNA was isolated using Trizol® Reagent (Invitrogen, Carlsbad, CA), according to the manufacturer's instruction and digested with DNase I (Promega, Madison, WI). cDNA was subsequently generated with oligo dT20) primers (Invitrogen) and Superscript ™ III Reverse Transcriptase (Invitrogen), according to the manufacturer's protocol.
