avian myeloblastosis virus  (TaKaRa)

Journal: Acta Pharmacologica Sinica

Article Title: Exendin-4 ameliorates oxidized-LDL-induced inhibition of macrophage migration in vitro via the NF-κB pathway

doi: 10.1038/aps.2013.128

Figure Lengend Snippet: RNA extraction and semi-quantitative RT-PCR

Article Snippet: TaKaRa RNA PCR Kit (AMV; version 3.0) was acquired from Takara Biotechnology Co, Ltd (Dalian, China).

Techniques: RNA Extraction, Quantitative RT-PCR

Journal: Oncology Letters

Article Title: Regulatory effect of Act1 on the BAFF pathway in B-cell malignancy

doi: 10.3892/ol.2019.10047

Figure Lengend Snippet: pTT5-Act1 expression vector construction. (A) Total RNA extraction from Raji cells. (B) A gel extraction kit was used to purify cDNA fragments of Act1 and pTT5 following dual restriction enzyme digestion (M, Marker; 1, 2 and 3, polymerase chain reaction products of Act1). (C) Construction of the pTT5-Act1 expression vector (M, Marker; 1, 2, 3 and 5, recombinant plasmid of pTT5-Act1). (D) A positive clone of pTT5-Act1 was verified using dual digestion (M, Marker; 1 and 2, sequences of pTT5 and Act1). Act1, nuclear factor-κB Activator 1.

Article Snippet: RNA isolation and reverse transcription-quantittive polymerase chain reaction (RT-qPCR) Total RNA was extracted from cells using the RNA PCR kit (AMV), version 3.0 (Takara Bio, Inc., Otsu, Japan) following the manufacturer's protocols.

Techniques: Expressing, Plasmid Preparation, RNA Extraction, Gel Extraction, Marker, Polymerase Chain Reaction, Recombinant

Journal: BMC Microbiology

Article Title: A gonococcal homologue of meningococcal ?-glutamyl transpeptidase gene is a new type of bacterial pseudogene that is transcriptionally active but phenotypically silent

doi: 10.1186/1471-2180-5-56

Figure Lengend Snippet: Transcriptional expression of the ggh genes. A. Dot blot analysis using the meningococcal ggt gene as a probe. One to 16 micrograms of RNAs isolated from H44/76, HT1089 (H44/76 Δggt::spc ) [39], ATCC49226, NIID54, NIID103 and NIID106 were subjected to this analysis. B. RT-PCR to detect the transcripts of the gonococcal ggh genes. The schematic figure in the box depicts the position of the primers used in this experiment (see Table 1). RT-PCR was performed without reverse transcriptase (RTase) (lanes 1 to 5) or with RTase (lanes 6 to 10). Lanes 1 and 6, H44/76 ( N. meningitidis ); lanes 2 and 7, ATCC49226 ( N. gonorrhoeae ); lanes 3 and 8, NIID54 ( N. gonorrhoeae ); lanes 4 and 9, NIID103 ( N. gonorrhoeae ); lanes 5 and 10, NIID106 ( N. gonorrhoeae ). The marker in the left-most lane is φ X174 DNA digested with Hae III. Primer sets used for RT-PCR are shown on the left side, and the corresponding PCR products are indicated by arrows on the right side. C. Primer extension analysis to detect the transcriptional start point of the ggt and ggh genes. Total RNA extracted from H44/76 ( N. meningitidis ), ATCC49226 and NIID106 ( N. gonorrhoeae ) was used for the primer extension with AMV reverse transcriptase XL and biotin-labeled oligonucleotide ggt-ext-2. The arrow on the right side indicates the transcriptional start site. D. Alignment of the nucleotide sequences of the upstream regions of the ggt and ggh genes. The sequence data have been deposited in the DDBJ/EMBL/GenBank Databases under the following Accession Numbers: N. meningitidis strains H44/76 [DDBJ: AB193252 ], H114/90 [DDBJ: AB193253 ], N. gonorrhoeae strains ATCC49226 [DDBJ: AB193254 ], NIID54 [DDBJ: AB193255 ], NIID103 [DDBJ: AB193296 ], NIID106 [DDBJ: AB193256 ]. An identical nucleotide is represented as *. The transcriptional start site is shown in bold as +1. The putative -35, -10 elements and Shine-Dalgarno sequence (SD) are depicted in the box, and the ideal -35 and -10 nucleotide sequences are shown above the boxes. The previously predicted start codon (ATG) [25] and newly predicted start codon (GTG) of the meningococcal {\it ggt} gene are underlined. The amino acid sequence deduced from the putative start codon GTG (shown in bold) in the meningococcal {\it ggt} gene is also shown under the corresponding nucleotide sequences.

Article Snippet: The hybridized RNA-DNA probe was treated with 35 units of AMV reverse transcriptase (RTase) XL (Takara Bio) in a reaction mixture (250 μM dNTPs, 1 × AMV reverse transcriptase XL buffer) at 37°C for 30 min.

Techniques: Expressing, Dot Blot, Isolation, Reverse Transcription Polymerase Chain Reaction, Marker, Polymerase Chain Reaction, Labeling, Sequencing

Journal: Immunology letters

Article Title: Modeling Sj?gren's Syndrome with Id3 Conditional Knockout Mice

doi: 10.1016/j.imlet.2010.09.009

Figure Lengend Snippet: Conditional deletion of the Id3 gene in developing thymocytes (A) Generation of the Id3 conditional deletion allele. Open box: exon. Hatch box: selection markers. Solid arrowhead: loxP. Open arrowhead: FRT. Arrows: primers. (B) Representative genotyping results of the Id3 f allele. DNA samples were prepared from mouse toes. 4 primers were used: Id3-2 and Id3-3 for the loxP (1.25 kb) and the wild type (1.15 kb) Id3 allele, Tek-F and Tek-R for the LckCre transgene (0.48 kb). M: 1 kb plus DNA ladder. (C) Deletion of Id3 in thymocytes sorted from 7-week-old mice. Thymocytes were separated into the DN (CD4 − CD8 − ) and DP/SP (CD4 + CD8 + , CD4 + CD8 − , and CD4 − CD8 + ) groups by magnetic beads. For the upper panel, the floxed and deletion alleles were determined by competitive PCR using three primers, Id3-2, Id3ckoF and Id3ckoB shown in (A). It produced a 1.07-kb band for the Id3 f allele and a 1.34-kb band for the deleted allele. For the bottom panel, only two primers (Id3ckoF and Id3ckoB) were used to detect the floxed allele. (D) Real-time quantitative PCR analysis of Id3 expression. RNA was prepared from total thymocytes of 7-week old Id3 f/f (open bar, n=4) and Id3 f/f ;LckCre (filled bar, n=3) mice. **P=0.006. The graphed results are means with SEM.

Article Snippet: DNase I treatment and reverse transcription were carried out with TaKaRa RNA PCR Kit (AMV) according to the manufacturer’s instructions.

Techniques: Selection, Mouse Assay, Magnetic Beads, Polymerase Chain Reaction, Produced, Real-time Polymerase Chain Reaction, Expressing