Notes: Total RNA was isolated from immature barley anthers with the RNAgents^® Total RNA Isolation System. Total RNA concentration was determined by spectrophotometric analysis. Next, the authors used the PolyATtract^® mRNA Isolation System to isolate poly (A)+ RNA from the total RNA samples. The purified poly (A)+ RNA was used as a template in RT-PCR using the Access RT-PCR System to analyze expression of BEN1 and Bnuc1. (2847)

Notes: In this study, the Wizard^® SV Genomic DNA Purification System was used to purify genomic DNA from Hepatitis C (HCV)-infected PH5CH8 hepatic cells. The purified genomic DNA was used in PCR to detect HCV-infected cells.  (3016)

Notes: The authors evaluated criteria for using STR analysis to resolve immigration cases by examining 156 one-parent child combinations and 105 two-parent child combinations from 28 families. STR analyses were performed in duplicate using the SMG Plus^® kit and a combination of CTTv and FFFL Multiplexes. The authors concluded that the combination of these kits, which analyze a total of 21 STR loci, can be used to successfully resolve immigration casework. However, in 9% of the one-parent cases their criteria of a parental index of 1.000 or more was not met. Additional testing using the PowerPlex^® 16 System reduced this from 9% to 1 out of 156 indices. (3858)

Notes: These authors characterized cytochrome P450 expression in mouse ovaries treated with the ovotoxin 4-vinylcyclohexene. Total mouse ovary RNA was reverse transcribed using the Reverse Transcription System and random primers, followed by quantitative PCR using primers specific for CYP2E1, 2A and 2B. (3442)

Notes: Total RNA was obtained from non-small cell lung cancer cell lines and the breast cancer resistance protein message was amplified using real-time quantitative RT-PCR. RT-PCR products were subcloned into the pGEM^®-T Easy Vector. (2714)

Notes: In this paper, the SV Total RNA Isolation System was used to isolate total RNA from human osteoblast HOBIT cells. One microgram of the purified DNA was used in RT-PCR with a 20mer oligo(dT) and gene-specific primers. (2850)

Notes: The authors used ISSR PCR to quantitate genomic instability using matched tumor (OSCC) and normal oral squamous cell samples. The inter-repeat region bands of similar molecular size that were altered in more than one case of OSCC were reamplified, gel purified and cloned into the pGEM T Vector for sequencing. (2635)

Notes: Researchers performed a 5´ RACE analysis on the human CCR8 and mouse CX3CR1 transcripts.  PCR-amplified products from the reactions were cloned in to the pGEM^®-T Vector for further analysis.  Also, the human CCR8 and mouse CX3CR1 promoters were cloned into the pGL3-Basic Vector and transfected into THP1 and Jurkat cells. For transfections, 15 x 10⁶ cells in 750μl were used with 15μg of construct in electroporation reactions. Transfectants were grown for 48 hours and analyzed with the Bright-Glo™ Luciferase Assay System. A fourfold increase in activity was observed compared to the pGL3-Basic Vector alone. (2729)

Notes: ImProm-II™ Reverse Transcriptase was used in real time RT-PCR to measure the ratio of Bax to Bcl-2 in immortalized rat proximal tubular cells (IRPTCs) cultured under normoxic or hypoxic conditions. The researchers used 1μg of total RNA in the reverse transcription reaction. Qiagen’s QuantiTest CYBR Green PCT Kit was used to quantify PCR products. Promega’s terminal deoxynucleotidyl transferase (TdT) was also used for TdT-mediated dUTP nick end labeling (TUNEL) assays on the cells. The TUNEL-stained cells were analyzed by FACS analysis. Data from these experiments was expressed as percent apoptotic cells.  (2849)

Notes: The SV Total RNA Isolation System was used to isolate total RNA from human placenta. The total RNA was used in RT-PCR to specifically amplify human FP prostanoid receptor 1 cDNAs. The amplified cDNAs were cloned and used in mutagenesis studies. (2852)

Notes: Researchers used M-MLV Reverse Transcriptase, RNase H Minus, Point Mutant, and random hexamers to reverse transcribe total RNA from various rat tissues. The cDNA was then used in real-time PCR reactions with SYBR Green dye and primers specific to Krim-1 or GAPDH sequences.  For reverse transcription, 200 units of M-MLV were used in each 25μl reaction.  Data from real-time PCR was normalized to GAPDH levels and expressed as relative expression levels in various tissues.  (2792)

Notes: The Wizard^® Genomic DNA Purification Kit was used to purify genomic DNA from various Streptococcal strains (Streptococcus sanguinis, S. oralis, S. gordonii, S. cristatus, S. mitis, S. parasanguinis, S. pneumoniae, S. mutans, S. salivarius, S. sobrinus, S. ratti, and S. vestibularis), Lactobacillus acidophilus, Actinomyces naeslundii, and E. coli JM109. The genomic DNA was diluted to 50μg/ml and used in Arbitrarily Primed PCR (AP-PCR) experiments to identify common amplimers from clinical Streptococcus sanguinis samples. The authors were able to identify a specific probe that may be helpful in identification of  S. sanguinis clinical isolates. (2836)

Notes: The Wizard^® SV Genomic DNA Purification System was used to purify DNA from cultured mammospheres. The mammosphere genomic DNA samples were then used in PCR reactions to amplify Short Tandem Repeats for genetic analysis. (2683)

Notes: The swarming behavior of the symbiotic-pathogenic bacterium Xenorhabdus nematophila is examined in this work. To investigate changes in gene expression in different strains at various times, the researchers utilized the AccessQuick™ RT-PCR System to measure the relative amount of gene product. To ensure that the assay was linear, different cycle numbers were used for each gene target ranging from 15 to 21 cycles of amplification. The RT-PCR used approximately 600ng of total RNA that was treated with RQ1 RNase-Free DNase before use.  (2745)

Notes: Terminal Deoxynucleotidyl Transferase was used to biotin end-label PCR products generated from adaptor-ligated genomic DNA fragment templates. The labeled probes were then hybridized to microarrays spotted with SNP alleles. The Terminal Deoxynucleotidyl Transferase reaction was performed for 4 hours at 37°C using 15-20 units TdT, TdT reaction buffer and 18μM biotin-ddATP. (2758)

Notes: Genomic DNA was isolated from Madurella mycetomatis samples.  A modified  Wizard^® Genomic DNA Purification Kit protocol was used for the isolations.  Mycelia were sonicated and ground in liquid nitrogen before the addition of Nuclei Lysis Solution. The purified DNA was used in PCR.  (3049)

Notes: The SV Total RNA Isolation System was used to isolate total RNA from Alteromonas cultures. Five micrograms of the total RNA was used in reverse transcription reactions with M-MLV Reverse Transcriptase, RNase H Minus. cDNA chiB (chitinase B) products from the reaction were quantified by real-time PCR with SYBR green dye. (3023)

Notes: The pGEM^®-T Easy Vector was used to subclone products of a 5´ RACE reaction. A promoter construct, assembled in the pGL3 Basic Vector, was co-transfected with a zebrafish interferon expression vector in the ZF4 zebrafish embryo fibroblast cell line using the TransFast™ Reagent (details provided). Luciferase levels were examined with the BrightGlo™ Luciferase Assay Reagent. Induction of zebrafish mRNA was also examined in zebrafish liver cells (ZFL) following treatment with the known interferon inducer, poly(I)-poly(C). RNA was extracted and reverse transcribed using ImProm-II™ Reverse Transcriptase. The resulting cDNA was used for quantitative, real-time RT-PCR with a SYBR green-based assay. (2627)

Notes: Researchers used the Wizard^® SV Gel & PCR Clean-Up System to purify rat c-fos RT-PCR products that were then quantified by A₂₆₀ absorbance. These products were used as standards for quantitative PCR reactions. The researchers used an Applied Biosystems Prism 7000 sequence detection system to perform the 50μl quantitative PCR reactions. (2678)

Notes: A region of the ORF from the APC gene from colorectal cancer and normal patients was isolated and amplified by PCR. The primers used incorporated transcription and translation sequence elements as well as N- and C-terminal FLAG and HA tags respectively. The PCR products were used as templates in TNT^® T7 Quick for PCR transcription/translation reactions.  Four different colored BODIPY-fluor-labeled lysines were added to the reactions to label the proteins in the TNT^® T7 Quick for PCR transcription/translation reactions. A subtractive precipitation strategy was used to purify full-length and truncated APC proteins based on the presences or absence of FLAG and HA tags. The partially purified proteins were run on SDS-PAGE gels and imaged with a Typhoon 9700 instrument to detect truncations.    (3026)

Notes: The Wizard^® SV Gel and PCR Clean-Up System was used to purify a 280 base-pair PCR product, which was then directly sequenced. (2791)

Notes: NF-E2-related factor-2 (NRF2) upregulates transcription of antioxidant response element (ARE)-driven genes. The authors examined the sensitivity of Nrf-2^–/^– primary mouse neurons to oxidative stress-induced apoptosis using the mitochondrial toxins 1-methyl-4-phenyl-1,2,5,6-tetrahydropyridine (MPP⁺) and rotenone. Primary cultures were >090% neurons, as judged by immunostaining with the Anti-βIII Tubulin mAb and other neuron-specific antibodies. Neuronal cytotoxicity in the presence of MPP⁺ and rotenone was monitored using the CellTiter 96^® AQueous Non-Radioactive Cell Proliferation Assay and TUNEL assays. The level of expression of the antioxidant-responsive gene quinone oxidoreductase was measured by mRNA quantitation using RT-PCR, enzyme activity assays and immunocytochemistry. The Reverse Transcription System was used to synthesize cDNA for subsequent amplification by PCR. (3570)

Notes: pGEM^®-T Easy Vector was used to clone PCR products.  The CytoTox 96^® Non-Radioactive Cytotoxicity Assay  was used to determine specific cytotoxicity of human dendritic cells that were transduced with recombinant adenoviruses containing the gene encoding a fusion protein of granulocyte-macrophage colony stimulating factor and carbonic anhydrase IX. (2674)

Notes: Promega T4 Polynucleotide Kinase was used to end-label an oligo representing a NF-kB binding sequence element with [γ-³²P] ATP (7,000 Ci/mmol).  Specific primers to IFN-β and IκB-α messenger RNA were used in RT-PCR to generate products for cloning into the pGEM^®-T Easy Vector. The resulting plasmids were linearized with Nco I and Spe I, respectively, and used as templates for in vitro transcription using the Riboprobe^® System to generate probes for use in an RNase protection assay.  The antisense probes were labeled with [α-³²P]CTP (800 Ci/mmol) in the Riboprobe^® reactions.  RNase ONE™ Ribonuclease was also used in this study.  (3197)

Notes: The ImProm-II™ Reverse Transcriptase was used to amplify the protein coding region of a 2,264 base mouse MT-1 gene mRNA and GAPDH from mouse total RNA.  RT-PCR was accomplished with Pfu DNA polymerase.  The RT-PCR product was used to make a probe for Northern analysis. (2609)