Journal: Neuroscience letters

Article Title: Involvement of GluR2 and GluR3 subunit C-termini in the trigeminal spinal subnucleus caudalis and C1-C2 neurons in trigeminal neuropathic pain

doi: 10.1016/j.neulet.2010.12.060

Figure Lengend Snippet: (A) Genotype in each generation of mutant mice confirmed by PCR analysis on isolated genomic DNA. PCR was performed on the isolated genomic DNA using taq DNA polymerase and primer sets (#1 and #2 are for wild-type and GluR2 delta7 KI, #3 and #4 are for

Article Snippet: PCR was performed on the isolated genomic DNA using taq DNA polymerase (TaKaRa Ex Taq™, Takara, Otsu, Japan) and primer sets (primer #21–23 for GluR2 delta7: #21, 5′-ACA GAG GAA GGT AGT GGA AGG GAG-3′; #22, 5′-CTT GGT TTG GTT GTT GGT CAT AGC-3′; #23, 5′-CTA GTG AAC CTC TTC GAG GGA C-3′; primer #31–33 for GluR3 delta7: #31, 5′-CCA ATA CTC CAC AGG GGC AAT TTA TC-3′; #32, 5′-CCG TTG ACT GTT TTG AAT CTC ACA CC-3′; #33, 5′-CTA GTG AAC CTC TTC GAG GGA C-3′).

Techniques: Mutagenesis, Mouse Assay, Polymerase Chain Reaction, Isolation

Journal: Journal of Virology

Article Title: Mutational Analysis, Using a Full-Length Rubella Virus cDNA Clone, of Rubella Virus E1 Transmembrane and Cytoplasmic Domains Required for Virus Release

doi:

Figure Lengend Snippet: Construction of RV M33 virus full-length cDNA clone. The numbers on the viral genome scale refer to the distance from the 5′ end in kilobases. Six DNA fragments were amplified by proofreading TaKaRa Taq DNA polymerase with the requisite primers as described in Materials and Methods. The amplified DNA fragments were ligated into a full-length cDNA representing the whole viral genome by using the restriction sites indicated above the genome. The full-length cDNA was cut with Eco RI and Hin dIII and inserted into a modified pBR322 plasmid that had been cut with the same enzymes to obtain the full-length cDNA clone pBRM33.

Article Snippet: PCR amplifications were performed in separate reactions containing cDNA, 1 pmol of primers, 0.4 mM deoxynucleoside triphosphates, 10% dimethyl sulfoxide, and proofreading TaKaRa Taq polymerase (Takara Shuzo Co., Ltd.) in a buffer provided by the manufacturer.

Techniques: Amplification, Modification, Plasmid Preparation

Journal: Journal of Bacteriology

Article Title: Viability of an Escherichia coli pgsA Null Mutant Lacking Detectable Phosphatidylglycerol and Cardiolipin

doi:

Figure Lengend Snippet: PCR analysis of the pgsA alleles of E. coli pgsA mutants. PCR products amplified with Ex Taq DNA polymerase were subjected to 1.2% agarose gel electrophoresis. Lanes 1, W3110; lanes 2, S301; lanes 3, MDL12; lanes 4 to 6, independent clones of the transductants (S330). Primer pairs FPPG5-ASFPPG1 (a) and FPPG5-ASFPPG3 (b) were used. The design and sequences of the primers are described in Materials and Methods. With the former primer pair, the wild-type pgsA allele and the pgsA :: kan allele gave products of 0.71 and 1.9 kbp, respectively. With the latter primer pair, the wild-type pgsA and the pgsA :: kan alleles gave products of 0.5 and 1.7 kbp, respectively. A DNA fragment of ca. 1.5 kbp which appeared in MDL12 (panel b, lane 3) may be the product of a false annealing of the antisense primer with a site in lacZ ′ fused to pgsA ). The molecular size markers included (two left lanes of each gel) were λ- Hin dIII digest (23.1, 9.4, 6.6, 4.4, 2.3, 2.0, and 0.56 kbp) and λ- Eco T14 I digest (19.3, 7.7, 6.2, 4.3, 3.5, 2.7, 1.9, 1.5, 0.93, and 0.42 kbp).

Article Snippet: DNA from fresh colonies of the strains to be examined was used as a template for PCR amplification with Ex Taq DNA polymerase (Takara, Tokyo, Japan).

Techniques: Polymerase Chain Reaction, Amplification, Agarose Gel Electrophoresis, Clone Assay

Journal: Frontiers in Microbiology

Article Title: A Multiplex PCR Detection Assay for the Identification of Clinically Relevant Anaplasma Species in Field Blood Samples

doi: 10.3389/fmicb.2020.00606

Figure Lengend Snippet: Optimization of the multiplex PCR components. (A) Annealing temperature gradients, M: DL2000 marker. Lanes 1–8: 64, 63, 62, 61, 60, 59, 58, and 57°C, respectively. (B) Dose of Anaplasma primers, lane M: DL2000 marker; Lanes 1–7: 0.08, 0.16, 0.24, 0.32, 0.4, 0.48, and 0.56 μM, respectively; Lane N: negative control. (C) La Taq DNA polymerase. M: DL2000 marker; Lanes 1–7: 0.75, 1.0, 1.25, 1.5, 1.75, 2, and 2.25 U, respectively; Lane N: negative control.

Article Snippet: Optimization of the Multiplex PCR After optimization, the optimum multiplex PCR assay was performed in a final volume of 25 μL, containing 2.5 μL of 10× PCR La buffer, 4 μL of dNTPs at 2.5 mM, 1.25 U of La Taq DNA polymerase, 0.32 μM of each primer, and 2 μL of the DNA template.

Techniques: Multiplex Assay, Polymerase Chain Reaction, Marker, Negative Control