Notes: IEC-6 cells were seeded into 96-well plates and cultured in the presence of various concentrations of human IL-17. Cell proliferation was measured using the CellTiter 96^® AQ_ueous Non-Radioactive Cell Proliferation Assay. The pGL3 vector was used to clone a PCR-amplified piece of the 5' flanking region of the CINC gene, and the Luciferase Assay System was used to assess promoter activation. All transfections of IEC-6 cells used the pSV-β-galactosidase vector as a control for transfection efficiency. Total RNA for Northern blotting was isolated using the Poly ATtract System. (2510)

Notes: The pALTER^®-MAX Vector was used to create mutations causing three separate amino acid substitutions on the FKHR cDNA. The Reverse Transcriptase System was used for RT-PCR. (1088)

Notes: The pGEM^®-3Zf(+) Vector was used for routine subcloning and the resulting plasmid was used as a template for PCR. (0114)

Notes: PCR products were purified with the Wizard® PCR Preps DNA Purification System and directly sequenced with the SILVER SEQUENCE™ DNA Sequencing System. (1334)

Notes: Total RNA was isolated from 3-day-old mung bean hypocotyls using the SV Total RNA Isolation System. RT for RT-PCR was performed with the Reverse Transcription System. (0825)

Notes: Total RNA was isolated from 5–10 mouse retinas with the SV Total RNA Isolation System. The isolated RNA was used for Northern analysis and RT-PCR. (1174)

Notes: Total RNA was isolated from mouse eyes using the SV Total RNA Isolation System. Four to six eyes produced sufficient quantities of RNA for Northern analysis (at least three blots at 5µg per lane) and RT-PCR. (0749)

Notes: The SV Total RNA Isolation System was used to extract total RNA from 106 SK-Mel2 human melanoma cells. The isolated RNA was used for RT-PCR with the Access RT-PCR System. The results were used for semi-quantitative analysis. (0959)

Notes: The Wizard^® Genomic DNA Purification Kit was used to isolate genomic DNA from Vibrio harveyi. The isolated DNA was used for PCR. (0060)

Notes: The initial work was performed with a single-tube RT-PCR system from a company in which both the reverse transcriptase and polymerase were mixed together and thus could not be separated. Consequently, the authors could not produce the important no-reverse transcriptase control. To do such a control, the authors used the Access RT-PCR System, which allows such control, since both the Tfl DNA Polymerase and AMV Reverse Transcriptase are added separately to the reaction. (0925)

Notes: The Access RT-PCR System was used for analytical RT-PCR for a variety of targets. Amplification was performed within the linear range of PCR product formation and normalized to the beta-tubulin signal or rpS6 signal. (1169)

Notes: Plasmids were purified with either the Wizard^® Plus Minipreps DNA Purification System or the Wizard^® Plus Midipreps DNA Purifications System. PCR products were purified with the Wizard^® PCR Preps System. (0751)

Notes: Before RT-PCR reactions, the RNA template was treated with RQ1 RNase-free DNase. (0776)

Notes: A PCR product amplified from a mutant RB (retinoblastoma protein) cDNA was cloned into the pCI-neo Mammalian Expression Vector. In vivo cyclin-mediated phosphorylation was assayed by cotransfecting the RB expression plasmids with members of the cyclin D or cyclin E family into an RB (-/-) cell line. (0571)

Notes: Genomic DNA was purified from blood or cultured fibroblasts, by means of the Wizard^® Genomic DNA Purification Kit. Total RNA was isolated from cultured fibroblasts by use of the SV Total RNA Isolation System, and it was amplified by Access RT-PCR System by use of exonic primers. An expression vector containing an S-tag attached to the 5´ end of the FALDH cDNA was constructed using pCI-neo Mammalian Expression Vector. (0482)

Notes: The PolyATtract^® System 1000 was used to isolate poly A+ RNA directly from bovine retinas. The isolated RNA was used for RT-PCR amplification. (0369)

Notes: The RNAgents^® Total RNA Isolation System was used to isolated total RNA from Porphyromonas gingivalis cultures. The isolated RNA was used for RT-PCR. The Prime-a-Gene^® Labeling System was used to make probes for Southern analysis. (0366)

Notes: Genomic DNA was isolated from Nectria haematococca mycelia with the Wizard^® Genomic DNA Purification Kit using a slightly modified protocol.  Mycelia were ground in liquid nitrogen and lyophilized before lysing in a modified lysis buffer consisting of 50mM Tris/HCl, pH 7.5, 50 mM EDTA (pH 8.0), 3% SDS and 1% mercaptoethanol.  The isolated DNA was the template for PCR. (0158)

Notes: This paper describes a quick method for analyzing plant microsatellites, also referred to as Simple Sequence Repeats (SSRs), using adaptor-ligated genomic DNA, PCR, and the Streptavidin MagneSphere^® Paramagnetic Particles. Briefly, 200μg of Streptavidin MagneSphere^® Paramagnetic Particles (pre-equilibrated with 6X SSC) were added to 100ng of a PCR product annealed to a biotinylated-repeat oligo. The captured fragments were washed, size-fractionated and used in PCR with AP-11 primers. (3050)

Notes: cDNAs for the receptor genes were obtained by RT-PCR using the Access RT-PCR System. (0569)

Notes: The size of PCR products were determined in relation to the PCR Markers. (1061)

Notes: pGEM^®-T Easy Vector Systems (2068)

Notes: A minigene was constructed with five exons and four introns from exons 5-9 of the CYP27 gene. The 2111bp product was generated with primers containing a start codon and a stop codon. The product was directly cloned into the pTARGET™ Mammalian expression vector and transfected into COS-1cells. RNA was isolated from these cells and RT-PCR was performed to amplify the spliced product. This assay could distinguish between normal splicing of the third intron and aberrant splicing of due to the Arg362Ser mutation. The proteins produced by the normal and mutant minigenes were also assessed by immunoblotting. (1333)

Notes: PCR products were purified by Wizard^® PCR Preps DNA Purification System and sequenced by use of the fmol^® DNA Cycle Sequencing Systems. (0247)

Notes: To confirm the G to A mutation caused alternative splicing, a minigene containing exon 6 with or without the G to A mutation was amplified. The 2111bp minigene was T/A cloned into the pTargeT™ Mammalian Expression Vector and transfected into COS cells. The RNA was isolated from the transfected cells and analyzed by RT-PCR. The G to A mutation resulted in a truncated RT-PCR product. To confirm the mutation would cause a truncated, inactive protein. A cDNA was constructed that would be the result of the alternative splicing and subcloned into the pTargeT™ Vector. COS cells expressing the mutant had no detectable sterol 27 hydroxylase activity. (1330)