Simultaneous In Vitro Protein Synthesis Using Solid‐Phase DNA Template
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N-Myristoyl transferase-mediated protein labelling in vivo
analysis. The straightforward synthesis of the chemical and molecular biological tools described should enable their use in a wide range of N-terminal labelling applications. Introduction Protein labelling using bioorthogonal ligation chemistry 15 55 Site -specific labelling of proteins with chemical or fluorescent
Sequencing of plant genomes a review
is acquired by the sequencing-by-synthesis principle. The bases are flowed sequentially across the device, and when there is complementation with the template, a pyrophosphate signal is generated and recorded by a charge-coupled device camera. Accordingly, the simultaneous sequencing of the entire genome in picoliter-sized plates occurs.
Rapid and Error-Free Site-Directed Mutagenesis by a PCR-Free
from dsDNA plasmid by using a pair of endonucleases (Nt.BbvCI and Nb.BbvCI).26 An inherent limitation is that the plasmid may contain multiple BbvCI nicking sites. In addition, multiple steps are involved in this approach, which is time-consuming. Some new techniques based on chemical solid-phase gene synthesis have also been developed for improved
Protein Engineering Methods and Applications
It is based on the use of recombinant DNA technology to change amino acid sequences. The first papers on protein engineer ing date back to early 1980ies: in a review by Ulmer (1983), the prospects for protein engineerin g, such as X-ray crys tallography, chemical DNA synthesis, computer modelling of protein structure and folding were discussed and
In Vivo Gene Transfer Using Sulfhydryl Cross-Linked PEG
in reversibly stabilized DNA condensates.12 16 In principle, the reducing environment of the endosome and cytosol can trigger the release of DNA intracellularly, which may account for the improved gene transfer efﬁciency observed for these reagents in vitro.14 16 Despite their demon-strated activity in vitro, none of the sulfhydryl
34th European Peptide Symposium and the 8th International
second-generation synthetic strategy of gm2-activator protein (gm2ap) analogues applicable to the preparation of a protein library pp i 031 evaluation of cys racemization during solid phase peptide synthesis under microwave irradiation pp i 033 investigating racemization in his couplings in spps pp i 034
Universal strategies for the DNA-encoding of libraries of
polymerase, template-dependent polymerization, unimolecular information recording and retrieval, in vitro selection The affinity-mediated selection of large libraries of DNa-encoded small molecules is increasingly being used to initiate drug discovery programs. We present universal methods for the encoding of such libraries using the chemical
RNA: packaged and protected by VLPs
and their protein capsids are T¼ 3 icosahedral nanoparticles with diameters of 30 nm, formed of 180 copies of the coat protein (CP).19 21 Qb VLPsarehollowwithpores1.4nmindiameter.20 Lau et al.22 and Fang 15 previously demonstrated pack-aging of non-viral RNA in Qb VLPs by co-expressing target RNA and CP from distinct plasmids in E. coli.
Molecules 2000, 5, 1259-1264 - MDPI
Figure 1: Schematic outline of mRNA-protein fusion synthesis. A double stranded DNA template library was in vitro transcribed to produce the mRNA template (a) which was annealed with a limiting amount of biotinylated puromycin-linker (b) and subsequently captured on neutravidin beads (c) After a washing step to remove unbound mRNA (d) the beads
Carlos L. Araya NIH Public Access Douglas M. Fowler Author
500 bp) from solid-phase arrays of DNA molecules simultaneously. Currently available platforms include Illumina, Roche/454, ABI/SOLiD, Polonator, Helicos, and Pacific Biosciences. Sequencing proceeds by synthesis, where base incorporations are monitored by fluorescence, or in the case of pyrosequencing, by luminance . The sample
Preface p. v
Detection of Mutations in DNA and RNA by Chemical Cleavage p. 685 Mutation Screening Using PCR-SSCP: Silver Staining and Isotopic Protocols p. 695 Detecting Point Mutations by Denaturing-Gradient Gel Electrophoresis p. 705 Analysis of Nucleotide Sequence Variation by Solid-Phase Minisequencing p. 717 The Amplification Refractory Mutation System
Rapid DNA protein cross-linking and strand scission by an
(Scheme SI) were prepared via solid phase chemical synthesis, and the 146-bp a-satellite DNA duplexes were constructed by ligating these with two longer biopolymers that were also chemi cally synthesized (Fig. SI). To ensure selective 5'-32P labeling of the strand containing 4, the DNA substrates were constructed
TechNote 102 Magnetic Microspheres
standard (non-protein coated) microsphere suspensions do not contain an antimicrobial agent. It is recommended that all suspensions be handled using aseptic technique. If possible, continuous rolling (e.g. 3-5 rpm on a cell culture roller) is recommended to keep microspheres in suspension, without generating foam
RNA multimerisation in the DNA packaging motor of
reasonable scope of RNA produced by in vitro transcrip-tion or chemical synthesis. As alternatives, two methods using the analytical ultracentrifuge have been employed. The ﬁrst method makes use of the difference in solvent densities for H 2O and D 2O-based buffers . It can be seen from equation (1) that the buoyant molecular weight, M
Nucleic-acid-binding properties of the C2-L1Tc nucleic acid
end of C2-L1Tc protein has a slight destabilization effect on a mismatched DNA duplex and shows a strong preference for single-stranded nucleic acids, such as C2-L1Tc. These results provide further insight into the essential properties of the C2-L1Tc protein as a NAC. Key words: double-stranded nucleic acid, long interspersed
RNA interference - Bio-Synthesis Inc
Unlike DNA, RNA possesses an additional hydroxyl group at the 2′ position of each ribose building block, which destabilizes RNA under the basic conditions generally present in DNA synthesis reactions. Hence, the most difficult step in RNA synthesis is the simultaneous protection of the 5′- and 2′-hydroxyl groups during solid-phase chemistry.
Entwicklung eines neuartigen PCR basierten Biochips
for DNA attachment on glass and efficient generation of solid-phase amplified DNA colonies. Nucleic Acids Res, 34, e22. Felbel, J., Bieber, I., Pipper, J. & Kohler, J. M. (2004) Investigations on the compatibility of chemically oxidized silicon (SiOx)-surfaces for applications towards chip-based polymerase chain reaction.
Advances in functional protein microarray technology
ging  to isolate protein complexes from cell extracts. Of these, protein separation by 2D-GE and subsequent identiﬁcation using MS have remained the two core technologies for large-scale proteomics. The 2D-GE method entails the separation of complex protein mix-tures by molecular charge in the ﬁrst dimension and by mass in the second
Publications by Astbury Centre Members 2007
121 Publications by Astbury Centre Members 2007 Allen, L. R. & Paci, E. (2007). Transition states for protein folding using molecular dynamics
Nanoparticle: An overview of preparation and characterization
elements for large-scale synthesis of organic and inorganic nanostructures with dimensions of 1 to 100 nm (Tomalia et al., 2004). They are compatible with organic structure such as DNA and can also be fabricated to metallic nanostructure and nanotubes or to possess an encapsulation capacity (Fu et al., 2007).
A ribozyme that ligates RNA to protein
RNA tags to proteins both in vitro and within bacterial cells, suggesting a simple way to tag a speciﬁc protein with ampliﬁable information. in vitro selection nucleic acid tag phosphoamide Tat TAR In vitro selection has been used in combination with rational design to identify and optimize numerous ribozymes and binding motifs (1 3).
Rapid DNA protein cross-linking and strand scission by an
DNA protein complexes, allowed for simultaneous quantifica-tion of intact DNA within core particles, DNA containing single strand breaks (SSBs) at the AP site, and DPCs. However, it does not distinguish DPCs containing a SSB from those in which the DNA is intact. To determine total strand scission regardless of
BEADS ABOVE THE RES T Magnetic Microsperes TechNote 102 9025 Technology Dr. Fishers, IN 46038 www.bangslabs.com [email protected] 317.570.7020 fax 371.570.7034 TechNote 102 Rev. #009, Active: 16/January/2017 Page 1 of 4
ROLE AND APPLICATIONS OF COMBINATORIAL CHEMISTRY IN MODERN
IN VITRO & IN SILICO ADME MODELS Potentially producing many more targets and personalized targets Screening up to 100,000 compounds a day for activity against a target protein Using a computer to predict activity Rapidly producing vast numbers of compounds Computer graphics & models help improve activity