Here, we describe a fast, simple method for constructing full-length cDNA libraries using SMART technology. This novel procedure uses the template-switching activity of Moloney murine leukemia virus (MMLV) reverse transcriptase to synthesize and anchor first-strand cDNA in one step. Following reverse transcription, three cycles of PCR are performed using a modified oligo(dT) primer and an anchor primer to enrich the cDNA population for full-length sequences. Starting with 1 microgram human skeletal muscle poly(A)+ RNA, a cDNA library was constructed that contained 3 x 10(6) independent clones with an average insert size of 2 kb. Sequence analysis of 172 randomly selected clones showed that 77% of cDNA clones corresponding to known genes contained intact open reading frames. The average length of complete open reading frames was 2.4 kb. Furthermore, 86% of the full-length clones retained longer 5' UTR sequences than the longest 5' end deposited in the GenBank database. cDNA libraries generated using this method will be useful for accelerating the collection of mRNA 5' end sequence information, which is currently very limited in GenBank.
Rabbits (n = 11) were fitted with large PMMA lenses to completely occlude the cornea and the anterior chamber lactate measured. The corneal epithelium was examined by light microscopy. The time period was 12 h. The results were compared with other authors' finding. There was a similarity of findings. The levels of lactate were found to have increased and the epithelium though edematous not stripped off. An individual response appeared to be the reason for the large variation in the individual findings.
Double layered metallic gratings have been investigated both theoretically and experimentally. The authors have reported that tunable magnetic resonance (MR) can be achieved by modulating the vertical chirped width dh which could be controlled conveniently in the common electron and/or ion beam microfabrications. The linear relationship between MR wavelength and dh has been reported. By introducing the difference of electric and magnetic penetration depth, an analytic formula deduced from a modified LC model has shown good agreement with the simulation results, and an effective width for trapezoidal sandwiched microstructures has been presented. Our results may provide an alternative choice for tunable MR and broad bandwidth of magnetic metamaterials.
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