A large number of sequenced plasmids in 7, 10, and 14 days libraries were in vitro transcripted (Riboprobe® in vitro Transcription Systems, Promega) and the RNA products were further reverse transcripted into the driver cDNA (Clontech PCR-Selected cDNA Subtraction Kit).
Stool samples were tested for the presence of RV using enzyme immunoassay, and a random subset of RV-positive samples was further genotyped using reverse transcriptase-polymerase chain reaction and reverse hybridization assay.
Fraction 1 was further purified by reversed-phase HPLC using THF H2O (50:50–80:20) as mobile phase to afford 1-O-palmitoyl-2-O-stearidonoyl-3-O-β-D-galactopyranosylglycerol (4) (Fig. 2) as an amorphous gum.
The fusion product of FIG and ROS was further confirmed by reverse-transcriptase-PCR (Figure 2C).
The GST-tag was removed using 3C protease and FnBR9,10 was further purified by reversed-phase HPLC.
PTEN exon 8 was further evaluated by reverse direction Sanger sequencing which also revealed no mutations (data not shown).
The tissue distribution of transgene expression was further assessed by reverse transcriptase polymerase chain reaction in six different tissues.
The synthesized peptide was further purified with reverse phase high performance liquid chromatography (RP-HPLC) using a 0-80% gradient Buffer B (99.9% acetonitrile, 0.1% trifluoroacetic acid), measuring absorbance at 214 nm.
The expression of selected genes was further analyzed by reverse-transcription PCR.
Crude extract of enterocin was further purified by reverse-phase HPLC Hewlett Packardd 1050, Agilent Technologies, Palo Alto, CA, USA).
