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)

Notes: The authors used RQ1 RNase-Free DNase to remove contaminating genomic DNA from total RNA samples. Cu, Zn, and MN SOD mRNA sequences were amplified from hippocampal neuronal RNA using the Access RT-PCR System. (2218)

Notes: PCR was used to amplify the VHL coding and promoter region in six fragments from genomic DNA, and SSCP analysis was performed to detect intragenic mutations. Fragments producing an aberrant SSCP band were purified using the Wizard^® PCR Preps DNA Purification System and subsequentially sequenced. (0176)

Notes: Total RNA was isolated from rat aorta and iliac arteries with the RNAgents® Total RNA Isolation System. The isolated RNA was used for RT-PCR. (1379)

Notes: The Access RT-PCR System was used to determine the presence of hepatitis C virus RNA in chimpanzee serum. (1465)

Notes: The pGEM® -T Easy Vector Systems, Wizard® PCR Preps DNA Purification System and Wizard® Plus Minipreps DNA Purification Systems were used in this study. (1959)

Notes: The S100A2 protein cDNA was subcloned into the pGEMEX^®-2 Vector and expressed in BL21(DE3)pLysS E.coli cells. The protein was purified by a referenced method. (1173)

Notes: The Access RT-PCR System was used to compare the abundance of GnT-IV mRNA in various bovine tissues. (0694)

Notes: Luciferase reporter constructs were cotransfected into 3T3-L1 preadipocytes with the pRL-CMV Vector at a 25:1 ratio and luciferase activities were determined with the Dual-Luciferase^® Reporter Assay System. RT-PCR was accomplished with AMV Reverse Transcriptase and Taq DNA Polymerase. RNase Protection Assay were performed with the aid of the RNase ONE™ Ribonuclease. (0988)

Notes: In this paper, Tfl DNA polymerase was used for two-step RT-PCR. The amplified fragments were cloned into the pGEM®-T Vector. Promega's Terminal Deoxynucleotidyl Transferase (TdT) was used to end label specific oligos for screening a frog brain cDNA library. T7 and T3 RNA polymerases were used to make digoxigenin labeled riboprobes for in situ hybridization. (1281)

Notes: The pGEM®-T Easy Vector was used to clone PCR products amplified from the prophage inserts of various lysogenic Lactococcus strains. (0193)

Notes: The Wizard® Genomic DNA Purification Kit was used to purify genomic DNA from Haemophilus influenzae. The isolated DNA was used for amplification. (1310)

Notes: For the RT-PCR experiment, total RNA isolated from fly heads was reverse transcribed by Promega AMV-RT and then the cDNA was amplified by Promega Taq DNA polymerase. (0574)

Notes: Total RNA was reverse-transcribed with random hexamer primers and AMV Reverse Transcriptase and then amplified for sequence analysis by nested PCR. Poly(A)+ RNA was purified with the PolyATtract^® mRNA Isolation System. (0382)

Notes: Genomic DNA was isolated from Methanococcus jannaschii using the Wizard^® Genomic DNA Purification Kit. The isolated DNA was used for PCR and eventually expression of the FtsZ protein. (0746)

Notes: The authors used the PolyATtract^® mRNA Isolation System IV to isolate mRNA from total RNA and the Wizard^® PCR Preps DNA Purification Resin to purify a first-strand cDNA synthesis product.  TNT^® T7 Coupled in vitro Transcription/Translation Reticulocyte Lysate System was used to synthesize radiolabeled DNaseY. And to test for signal peptide function they added Canine Pancreatic Microsomal Membranes (CMMs). (0779)

Notes: The authors compared a commercially available enterovirus-specific RT-PCR kit with a kit of their own design. Their 'in-house' RT-PCR system used the Access RT-PCR System to amplify enterovirus sequences from cerebral spinal fluid extracts followed by nested-PCR for final amplification of the target. Both the commercial system and the 'in-house' system were adequate for amplification of enterovirus sequences from CSF of infected individuals. Other, more time-consuming assays were performed to verify the results from the RT-PCR tests. (0545)

Notes: Quantitative RT-PCR was carried out using 1µg total RNA from Arabidopsis using Access RT-PCR System. Samples were taken during the PCR reaction after 5, 10, 15, and 25 cycles and loaded onto agarose gels. Gels were treated for Southern blot hybridization, and filters were hybridized using the AtMYB2 cDNA. Linearity of signal strength was verified using phosphorimaging system quantifications. (1048)

Notes: The Access RT-PCR System was used to amplify two portions of the CHD1 gene product from ostrich poly A+ RNA. The amplification products were gel purified and sequenced. The sequence obtained was compared to that of the human and murine CHD1 and Chicken CHD1Z and CHD1W sequences. (1175)

Notes: The SV Total RNA Isolation System was used to isolate total RNA from normal skin, psoriatic skin, H-128 small cell lung carcinoma cells and peripheral blood mononuclear cells. The isolated RNA was used for RT-PCR performed with the Access RT-PCR System. (0107)

Notes: The fmol^® DNA Cycle Sequencing System was used in this study. (0679)

Notes: Bisulphite-treated DNA was purified using the Wizard® DNA Clean-Up System prior to PCR. The amplified DNA was then cleaned using a Wizard® PCR Preps DNA Purification System prior to cloning and the cloned DNAs were sequenced using the fmol® DNA Cycle Sequencing System. (0001)

Notes: The Wizard® Genomic DNA Purification Kit was used to isolate genomic DNA from 3ml S. cerevisiae cultures grown 18-20 hours in YPD broth. The isolated DNA was used for PCR amplification of a 3.5kb segment. The amplimer was purified using the Wizard® PCR Preps System. (0356)