A high yield, rapid and simple procedure is described for extracting RNA from mycobacteria and other micro-organisms refractory to disruption. The method yielded 20 microg RNA/109 Mycobacterium bovis BCG, more than 10 times greater than our previous method. Intact full length hsp 70 (dnaK) mRNA was detected by northern blotting and quantitated after heat shock by slot blot hybridisation.

Comparative genomics reveals a common theme of 20S proteasome and proteasome-activating nucleotidase genes dispersed throughout archaeal genomes yet arranged in conserved linkages with gene homologues of translation and/or transcription machineries. To provide biological evidence for these linkages as well as insight into proteasome operon organization, transcripts of the five proteasomal genes...

By screening a cDNA library constructed from aortic total RNA derived from Watanabe heritable hyperlipidemic (WHHL) rabbits by differential hybridization, we have obtained a cDNA encoding the kappa light chain of immunoglobulin. Northern blot analysis of total RNA prepared from aortas of WHHL and normal rabbits of various ages revealed that this light-chain mRNA accumulates gradually with age i...

Here we describe a northern blot procedure that allows the detection of endogenous RNAs as small as approximately 14 nt in total RNA extracts from bacteria. RNAs that small and as part of total bacterial RNA extracts usually escape detection by northern blotting. The approach combines LNA probes 5'-digoxigenin-endlabeled for non-radioactive probe detection with 1-ethyl-3-(3-dimethylaminopropyl)...

Rapid, quantitative Western blotting is a long-sought bioanalytical goal in the life sciences. To this end, we describe a Western blotting assay conducted in a single glass microchannel under purely electronic control. The μWestern blot is comprised of multiple steps: sample enrichment, protein sizing, protein immobilization (blotting), and in situ antibody probing. To validate the microfluidic...

Anatomical models have always been a mainstay of descriptive embryology [5]. As the training of embryologists grew in the late 1800s, so too did the need for large-scale teaching models. Embryo wax models, such as those made by Adolf Ziegler [6] and Gustav Born [7], were popular in the latter part of the nineteenth century and the early twentieth century as a way to visualize, in three dimensio...

SKG or BALB/c T cells (3.0 £ 10) were stimulated for 72 h with plate-bound anti-CD3 mAb (2C11) in the presence or absence of plate-bound anti-CD28 mAb in RPMI-1640 medium supplemented with 10% fetal calf serum and 50 mM 2-mercaptoethanol. Cells were also stimulated with TPA (1.4 ng ml) and ionomycin (0.14 mM). KJ1.26þ T cells from DO or DO.SKG mice were stimulated with ovalbumin (323–339) pepti...

RQRT-PCR technique was evaluated for its validity as an alternative to Northern blotting for quantification of plant gene expression in diseased tissues of Hevea. Reliable RT-PCR results could be obtained by co-amplification of housekeeping actin gene as the internal control along with the gene of interest. The product of interest was quantified relative to that of the internal control by measu...