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: Total and poly(A)+ RNA from treated and untreated 8701-BC breast cancer tumor cells were treated with RQ1 RNase-free DNase. The RNA samples were reverse transcribed using the M-MLV RNase H– point mutant reverse transcriptase and random primers to create cDNA used in downstream analyses. The cDNA from the reactions were used in PCR, differential display PCR, and semi-quantitative multiplex PCR reactions. In differential display PCR studies, the authors analyzed differentially displayed bands by staining 6% polyacrylamide gels with the SILVER SEQUENCE™ Staining Reagents. Differentially displayed bands were re-amplified and cloned using the pGEM®-T Easy Vector and high efficiency JM109 competent cells. (3020)

Notes: A full length cDNA representing the entire genome of Diaporthe RNA virus (DaRV) was successfully cloned into the pGEM^®-T Easy Vector. The resultant construct was named pDV3. A positive strand viral RNA was then synthesized from the Sal I linearized pDV3 template. Fungi transfected viral RNA were shown to have different morphological features when compared to untransfected fungi. (2757)

Notes: The cDNA for the IL-1F7b protein was amplified from a human spleen cDNA library and directly subcloned in the pGEM^®- T Easy Vector and sequenced. The clone was transferred to a bacterial expression vector with a purification tag. The expressed protein was used to make a polyclonal rabbit antibody and antibody purification column. The cDNA for the IL-1F7b was reamplified and directly subcloned into the pTARGET™ Mammalian Expression Vector. The cDNA was stably expressed in RAW264.7 mouse monocyte/macrophage cell line.  Lysates from the cells stably expressing IL-1F7b were used to test the antibodies by western blotting. Expressing cells were also tested by immunocytochemistry. (2598)

Notes: The authors used the pGEM-T vector to clone a novel gene identified through RT-PCR analysis of human endogenous retrovirus K (HERV-K). (2469)

Notes: The promoter region of human CYP19 was amplified from a lambda library and cloned into the pGEM^®-T Easy Vector for sequencing. Several mutants of this aromase cytochrome P450 promoter were created and cloned into the pGL3-Basic Vector. One microgram of the various reporter constructs along with 25ng pRL-null Vector (as an internal control) were transfected into HMEC-1 and MCF-7 cells in six-well plates. Luciferase levels were assayed 48 hours post-transfection using the Dual-Luciferase^® Reporter Assay System. To further characterize the CYP19 promoter, a 30-mer region with a GATA motif was added to 250ng HMEC-1 nuclear extract in the presence of oligonucleotide competitors or antibodies using the Gel Shift Binding 5X Buffer, then analyzed by gel electrophoresis. (3110)

Notes: Researchers PCR amplified and cloned the Trl promoter into the pGEM^®-T vector. The Trl promoter sequence, as well as the eve promoter sequence and/or a CMV promoter, were used to make luciferase reporter constructs using the pGL3-Basic Vector. Transient transfection experiments were performed with the promoter constructs and GAGA factor expression constructs. Relative luciferase activity from each experiment was measured and compared to that of a control.  (2775)

Notes: The pGEM^®-T Easy Vector was used to clone a 12.5 kb fragment containing human kidney specific chloride channel (CLC-KB) gene fused to an enhanced green fluorescence protein (EGFP) created in a long and accurate PCR (LA-PCR) reaction.  The resultant construct was further manipulated and used to make transgenic mice.  (3131)

Notes: The authors used the pGEM-T Easy Vector to clone fragments amplified from chick Notch 1, Delta 1, Ngn 1, and Ng 2 in order to generate digoxigenin labeled antisense riboprobes. (2588)

Notes: The Access RT-PCR System was used to generate nuclear gene fragments from Chlamydomonas reinhardtii for use as probes for RNA blot hybridization assays and microarrays. RT-PCR reaction products were cloned into the pGEM^®-T Easy Vector for sequence verification and to allow easier manipulation. Details of the generation of microarray slides printed on GAPII glass slides (Corning) are provided. (2694)

Notes: Total RNA was isolated from Arabidopsis tissues using the RNAgents^® Total RNA Isolation System. The RNA was further processed into the poly(A) fraction using the PolyATtract^® mRNA Isolation System. The isolated RNA was used in Northern blot analysis. Blots were probed with a ³²P-labeled RNA probe generated from a cDNA cloned into the pGEM^®-T Easy Vector using the T7 Riboprobe^® in vitro Transcription System. Bioluminescence from transgenic plants containing a firefly luciferase reporter was visualized by spraying the plants with a solution of 5mM Beetle Luciferin, Potassium Salt in 0.1% Triton^® X-100. (2570)

Notes: The pGEM^®-T Easy Vector was used to clone the 5.4kb open reading frame of the kinesin-related protein (KRMP1). The gene was later subcloned into fusion tagged expression vectors and used in point mutation analysis of the protein’s active site.  For some of these studies, the researchers used Promega Recombinant Human cdc2 Kinase as a substrate in in vitro phosphorylation experiments.  (3136)

Notes: Taq Polymerase was used to amplify mitochondrial sequences from various laser-capture microdissected rat DNA samples. A 15.7 kilobase PCR fragment was cloned into the pGEM^®-T Easy Vector. The resultant clone was used to identify sequence breakpoints. (3234)

Notes: These researchers created alkaline phosphatase-conjugated probes that were specific for the α-proteobacterium Rhodospirillum rubrum RubisCO gene, rbcL. These probes were used in Southern and Northern blot analysis of non-transformed and transformed tobacco rubrum plants using the AttoPhos® AP Fluorescent Substrate System, a Vistra Fluorimager™ and ImageQuant™ software. The AttoPhos® Substrate was also used to visualize immunoblots of various proteins in transformed and non-transformed plants. To confirm their results, the researchers PCR-amplified choroplast DNA and ligated the products into the pGEM®-T Easy Vector for BigDye® Sequencing. (2649)

Notes: Total RNA was isolated from Mycobacterium smegmatis for Northern blot analysis and RT-PCR. Transcripts for furA and katG were amplified by RT-PCR using Promega's MMLV Reverse Transcriptase and cloned into the pGEM^®-T Easy vector. Northern blot probes for furA and katG were synthesized by in vitro transcription from the pGEM^®-T Easy vector. (2310)

Notes: Poly(A)⁺ RNA was purified from total RNA by the PolyATtract^® mRNA Isolation System. PCR products were purified by agarose gel electrophoresis and were cloned into pGEM^®-T and pGEM^®-T Easy Vector. Genes were obtained by RT-PCR using the Access RT-PCR System. (1097)

Notes: The authors used the Tfx™-50 Reagent to transfect bovine aortic endothelial cells. They also used the pGEM^®-T Easy Vector System to clone PCR products. (1482)

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

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 authors used the pGEM^®-T Easy Vector System in their studies. (1966)

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

Notes: In this paper, a 538bp fragment was amplified and subcloned into the pGEM^®-T Easy Vector. (1483)

Notes: These authors used Promega's pGEM^®-T Easy Vector Systems, Taq DNA Polymerase and Tli DNA Polymerase in this paper. (2028)