Journal:

Article Title: Fast IPSCs in rat thalamic reticular nucleus require the GABA A receptor ? 1 subunit

doi: 10.1113/jphysiol.2006.106617

Figure Lengend Snippet: A and B, scatter plots of IPSC 90%-width versus peak current amplitude for two RTN cells recorded on the same day. Each point in A and B represents an individual IPSC. A is a recording from a cell dominated with slow and small IPSCs (○) and B, represents a recording dominated by brief and large IPSCs (). C, ensemble averaged IPSCs averaged from isolated spontaneous inhibitory events (selected on the basis of having arisen from an event-free baseline and showing complete decay to the same baseline) for the two cells represented in A (black, n = 258) and B (grey, n = 217). Each trace was best fitted to two exponentials (dotted line superimposed on each trace). Note: the cell dominated with fast IPSCs yielded a ensemble averaged IPSC with a larger A1 value than A2 whereas the opposite is true for the slower cell. D, ethidium bromide stained 8% polyacrylamide gel showing single-cell RT-PCR analysis from five RTN cells examined in one day of recording, two of the cells are represented in A (lane 3) and B (lane 7). Cells positive for the β1 subunit cDNA showed a solitary band at 162 bp after two rounds of amplification using nested primers (lanes 6 and 7, see methods). Lanes 1 and 2 are water and media controls that underwent reverse transcription along with individual cells in lanes 3 through 7; lane 8 is a positive control using 40 pg of total RNA; lanes 9 and 10 are water controls used in the master mix for each round of PCR amplification. The molecular weight marker in lane 11 is the MspI digest of pBR322. The size of each fragment is noted on the right. The parameters for β1 negative cells in lane 4: A1 = 9.80, τ1 = 13.75, A2 = 14.17, τ2 = 64.61, τDW = 43.82l; and lane 5: A1 = 12.94, τ1 = 12.94, A2 = 5.69, τ2 = 77.21, τDW = 29.63. The β1 positive cell in lane 6: A1 = 18.05, τ1 = 15.11, A2 = 4.15, τ2 = 74.44, τDW = 26.20.

Article Snippet: The molecular weight marker was the pBR322 DNA– Msp I digest (New England Biolabs, Ipswich, MA, USA).

Techniques: Isolation, Staining, Reverse Transcription Polymerase Chain Reaction, Amplification, Positive Control, Molecular Weight, Marker

Journal: Nucleic Acids Research

Article Title: A conserved 3′ extension in unusual group II introns is important for efficient second-step splicing

doi: 10.1093/nar/gkp186

Figure Lengend Snippet: In vivo splicing of unusual group II introns in B . thuringiensis kurstaki 4D1. RT-PCR was conducted on total RNA with primers located in the flanking exons. The gel picture shows the RT-PCR products of the spliced exons (names of the products related to each intron are given on top) and the corresponding negative controls run without reverse transcriptase (lanes marked with NC). Lane M, pBR322 DNA digested with MspI (New England Biolabs), as marker. Samples were separated on a 2.8% NuSieve GTG agarose gel (Cambrex).

Article Snippet: Lane M, pBR322 DNA digested with MspI (New England Biolabs), as marker.

Techniques: In Vivo, Reverse Transcription Polymerase Chain Reaction, Marker, Agarose Gel Electrophoresis

Journal:

Article Title: Systematic Mutagenesis of the Murine Gammaherpesvirus 68 M2 Protein Identifies Domains Important for Chronic Infection ▿

doi: 10.1128/JVI.02234-07

Figure Lengend Snippet: Construction of M2 mutant viruses and corresponding marker rescue viruses. (A) The MHV68 genome cloned as a BAC (2) was used to generate all M2 recombinant viruses by RecA-mediated recombination. For M2 mutant viruses exhibiting robust establishment-of-latency and/or reactivation-from-latency phenotypes, a marker rescue virus in which the wt M2 sequences were targeted to the M2 ORF by allelic exchange to restore the wt phenotype was generated. The DNA probe used for the Southern blot analyses contained sequence from the M2 ORF (genomic coordinates, bp 4031 to 4627). Relevant restriction sites are indicated. A, AluI; Bg, BglI; D, DdeI; N, NcoI; BU, BstUI; Bs, BssHII. (B and C) Southern blot analyses of recombinant viruses harboring mutations in the indicated PXXP motifs of M2, along with corresponding marker rescue viruses. (D) Southern blot analyses of recombinant viruses harboring mutations in the indicated tyrosine residues of M2 and their corresponding marker rescue viruses. (E) Southern blot analysis of M2.Stop(2). 32P-labeled molecular size standards (Lambda DNA-BstEII digest; New England Biolabs, Beverly, MA) were included in each Southern blot analysis. WT, wt MHV68; MR, marker rescue virus.

Article Snippet: 32 P-labeled molecular size standards (Lambda DNA-BstEII digest; New England Biolabs, Beverly, MA) were included in each Southern blot analysis.

Techniques: Mutagenesis, Marker, Clone Assay, Recombinant, Generated, Southern Blot, Sequencing, Labeling, Lambda DNA Preparation