Notes: Streptavidin MagneSphere^® Paramagnetic Particles (SA-PMPs) were used to capture biotinylated primer probe in a random access retrieval of genetic information by PCR (rargip) screening [ABE, K., (1992) Rapid isolation of desired sequences from lone linker PCR amplified cDNA mixtures: application to identification and recovery of expressed sequences in cloned genomic DNA. Mamm. Genome 2:252-259]. The pGEM^®-T Vector System and PolyATract^® mRNA Isolation System were also used. (0513)

Notes: PCR products were cloned into the PinPoint^® Xa1 T-Vector and expressed in JM109 cells. The cell extracts were electrophoresed and immunoblotted for the specific protein. (0626)

Notes: Total RNA was isolated from ~3 × 10⁶ RAW264.7 macrophages using the SV Total RNA Isolation System. The amount of recovered RNA was adjusted to 500ng/µl and stored at –20°C. One microgram of the RNA was used for RT-PCR analysis. (0066)

Notes: The PolyATtract^® System 1000 was used to isolate polyA+ RNA directly from peripheral blood leukocytes. A 30µl preparation of mRNA was isolated from 1.2 x10⁹ cells. Three microliters of the isolated RNA was used for RT-PCR amplification of the p53 message with the Access RT-PCR System. (1656)

Notes: cDNA synthesis was performed in a reverse transcription reaction containing single-stranded binding protein and RNasin^® Ribonuclease Inhibitor. In a protein truncation test (PTT), five overlapping segments amplified by RT-PCR of NF1 transcript were expressed by TNT^® Coupled Reticulocyte Lysate System. (1051)

Notes: The Access RT-PCR System was used to study expression of three ntn genes from TW3 cells grown in either 4-nitrotoluene, toluene or succinate. The pET-5a Vector (discontinued) was used to express the ntnC protein in E. coli strain BL21(DE3) pLysS. (0968)

Notes: The Access RT-PCR System was used to determine HPRT (hypoxanthine-guanine phosphoribosyltransferase) and iNOS (induced nitric oxide synthase) mRNA in salivary gland lysate of P. papatasi. The Griess Reagent System was used to measure the nitrite accumulation in macrophage culture media. (0191)

Notes: Poly-A+ RNA was isolated from rat cerebellar total RNA with the PolyATtract^® mRNA Isolation System and used for semi-quantitative RT-PCR. The targets of the RT-PCR were cloned with the pGEM^®-T Vector System and sequenced to show they quantitated the correct targets. (0671)

Notes: The SV Total RNA Isolation System was used to isolate total RNA from mouse fibroblasts. The isolated RNA was used for RT-PCR analysis with the Access RT-PCR System. (0474)

Notes: This paper discusses the inhibition of PCR by reverse transcriptases. The inhibitory effect was overcome by increasing template concentration to more than 10^5 to 10^6 copies or including T4 gene 32 protein during the reverse transcription phase of the reaction. The poly(A)+ RNA used in the study was isolated using the PolyATtract® mRNA Isolation System. (1357)

Notes: Chromosomal E.coli DNA was prepared with the Wizard^® Genomic DNA Purification Kit and used for PCR. The resulting PCR fragments were purified with the Wizard^® PCR Preps DNA Purification System and used for fluorescent sequencing. (0203)

Notes: Genomic analysis noted a single base change between wild type and mutant. To see if this base change was enough for alternative mRNA splicing, minigenes were constructed and subcloned into the pTargeT™ Vector. The mutant and wild-type minigenes were transfected into COS-1 cells. Forty-eight hours post-transfection total RNA was isolated and analyzed by RT-PCR. Transfectants with the mutant sequence did produce a different product due to alternative splicing. (1331)

Notes: Poly A+ RNA was isolated directly from PBS-perfused mouse lungs with the PolyATtract^® System 1000. The isolate poly A+ RNA was used for quantitative two-step RT-PCR using M-MLV RNase Hminus Reverse Transcriptase as well as RNasin^® Ribonuclease inhibitor for first step cDNA synthesis. (0821)

Notes: cDNA was generated by reverse transcription of RNA purified from lymphoblastoid cell lines. PCR amplification of the cDNA was used to introduce a 17bp consensus T7 promoter sequence and a mammalian translation-initiation sequence in frame with a unique hMLH1 or hMSH2 sequence. Resultant PCR products were translated in the TNT^® T7 Coupled Reticulocyte Lysate System. (1194)

Notes: ()

Notes: The Wizard^® Genomic DNA Purification Kit was used to isolate genomic DNA from transgenic mouse peripheral blood lymphocytes. The isolated genomic DNA was used for PCR analysis of the CD95 genotype. (0718)

Notes: The Access RT-PCR System was used for a semi-quantitative assessment of the quantity of Cu/Zn superoxide dismutase (Cu/Zn SOD) Mn superoxide dismutase (Mn SOD) mRNA levels. Primary embryonic hippocampal neurons were treated with Abeta(25-35) for zero to 25 hours. The level of the Cu/Zn SOD and Mn SOD mRNA was related as percent of control, untreated cultures. The PCR portion of the reaction was performed for 22 cycles to insure that the reaction was in the linear range. (0106)

Notes: PCR products were excised, eluted, and precipitated prior to sequencing with the fmol^® DNA Cycle Sequencing System. Promega's RNasin^® Ribonuclease Inhibitor and AMV Reverse Transcriptase were used in a reverse transcription assay. (2065)

Notes: The system was used to isolate total RNA from Jurkat cells. The isolated RNA was used for RT-PCR. The RT reaction was catalyzed by MMLV Reverse Transcriptase. (1662)

Notes: The authors used T4 RNA Ligase to circularize highly purified total mitochondrial tRNA prior to RT-PCR. (1083)

Notes: The pGEM®-5Zf(+) Vector was used for subcloning of a PCR fragment. The fragment was used to make a probe for RNase protection assays. The pGEM®-7Zf(+) Vector was used for routine subcloning of restriction fragments from the chitotriosidase gene. (1427)

Notes: The Access RT-PCR System was used to assess expression of the CIITA gene in fibroblast L and splenic B cells. The 700bp transcript could be detected with as little as 40ng of poly(A)+ RNA from the splenic B cells but undetectable in as much as 5µg of poly (A)+ RNA from the L cells. Both sources of RNA were readily amplified for a 510bp β-actin message from 5µg down to 40ng. (0657)

Notes: PCR products were purified by Wizard^® PCR Preps DNA Purification System and sequenced with the fmol^® DNA Cycle Sequencing System. (0576)

Notes: The Serine/Threonine Phosphatase Assay System was used to assess protein phosphatase 2A, 2B and 2C activity in nuclear extracts of Human umbilical vein cells (HUVEC) with or without lysophosphatidylcholine treatment. Gel shifts were also performed with nuclear extracts prepared from the HUVEC cells using the SP1 Consensus Oligonucleotide. Luciferase reporter studies were also performed in HUVEC cells using pGL3 Basic-derived vectors and the Luciferase Assay System. (1305)

Notes: The Tfx™-50 Reagent was used to stably transfect 293 cells. Successful stable transfection was judged by RT-PCR and functional assays. (1079)