purexpress in vitro transcription translation system (New England Biolabs)

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New England Biolabs purexpress in vitro transcription translation system
$Release rates and estimation of pulling forces. (A) The rate of release (kR) obtained from pulse-chase experiments (SI Appendix, Fig. S4 and Dataset S2), the fraction full-length protein (fFL) measured under standard experimental conditions (20-min incubation in <t>PURExpress</t> in the continuous presence of [35S]Met), and the pulling force F calculated using Eq. 1 (k0=3.0×10−4s−1, Δx‡=0.65nm). The constructs are from Kudva et al. (12) and are colored to match those in B and SI Appendix, Figs. S4 and S5. (B) F values calculated from Eq. 1 plotted against the standard fFL values, with constructs colored as in A. The least-squares fit line is indicated by the blue line, and the analytic relation Eq. 3 between F and fFL, assuming an average delay time Δt = 550 s (approximately equal to half the standard incubation time), is shown as a red curve.$
Purexpress In Vitro Transcription Translation System, supplied by New England Biolabs, used in various techniques. Bioz Stars score: 98/100, based on 677 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/spatial+transcriptomics+deconvolution+by+topic+modeling+(stride)+method/PURExpress/pmc07322005-285-1-9
Average 98 stars, based on 677 article reviews

purexpress in vitro transcription translation system - by Bioz Stars, 2026-10

98/100 stars

Images

1) Product Images from "Cotranslational folding cooperativity of contiguous domains of α-spectrin"

Article Title: Cotranslational folding cooperativity of contiguous domains of α-spectrin

Journal: Proceedings of the National Academy of Sciences of the United States of America

doi: 10.1073/pnas.1909683117

$Release rates and estimation of pulling forces. (A) The rate of release (kR) obtained from pulse-chase experiments (SI Appendix, Fig. S4 and Dataset S2), the fraction full-length protein (fFL) measured under standard experimental conditions (20-min incubation in PURExpress in the continuous presence of [35S]Met), and the pulling force F calculated using Eq. 1 (k0=3.0×10−4s−1, Δx‡=0.65nm). The constructs are from Kudva et al. (12) and are colored to match those in B and SI Appendix, Figs. S4 and S5. (B) F values calculated from Eq. 1 plotted against the standard fFL values, with constructs colored as in A. The least-squares fit line is indicated by the blue line, and the analytic relation Eq. 3 between F and fFL, assuming an average delay time Δt = 550 s (approximately equal to half the standard incubation time), is shown as a red curve.$
Figure Legend Snippet: Release rates and estimation of pulling forces. (A) The rate of release (kR) obtained from pulse-chase experiments (SI Appendix, Fig. S4 and Dataset S2), the fraction full-length protein (fFL) measured under standard experimental conditions (20-min incubation in PURExpress in the continuous presence of [35S]Met), and the pulling force F calculated using Eq. 1 (k0=3.0×10−4s−1, Δx‡=0.65nm). The constructs are from Kudva et al. (12) and are colored to match those in B and SI Appendix, Figs. S4 and S5. (B) F values calculated from Eq. 1 plotted against the standard fFL values, with constructs colored as in A. The least-squares fit line is indicated by the blue line, and the analytic relation Eq. 3 between F and fFL, assuming an average delay time Δt = 550 s (approximately equal to half the standard incubation time), is shown as a red curve.

Techniques Used: Pulse Chase, Incubation, Construct

other:

Article Title: tRNA-deacylase-directed discovery of biosynthetic pathways.
Article Snippet: All chemicals were purchased from Sigma-Aldrich and used without further purification.

Control:

Article Title: Intrinsic disorder in elicitin-like effectors: Molecular shields in the arms race of biotrophic pathogens
Article Snippet: .. In order to replace the DHFR control gene with the gene of interest, we digested the PURExpress ® DHFR control plasmid with the enzymes NdeI (NewEngland Biolabs, Cat. No R0111S) and XhoI (NewEngland Biolabs, Cat. No R0146S) followed by treatment with the T4 DNA polymerase (NewEngland Biolabs, Cat. No M0203S) to create blunt ends. ..

Plasmid Preparation:

In Vitro:

Article Title: Comparative characterization of Cas12f orthologs reveals mechanistic features underlying enhanced genome editing efficiency.
Article Snippet: .. First, 5 nM of nuclease-amplified DNA templates and 25 nM of gRNAamplified DNA templates, including one of the spacer sequences 5′-GTCGAGGCTTGCGACGTGGT-3′ (U67 spacer) or 5′-TGGAGA TATCTTGAACCTTG-3′ (U40 spacer), were expressed at 37 °C for 3 h with a PURExpress in vitro protein synthesis kit (NEB). .. Plasmid library DNA cleavage reactions were carried out by mixing 5 nM of the target library representing all possible 8N PAMs, a fivefold dilution of PURExpress expressions, 10 mM Tris-HCl pH 7.9, 10 mM MgCl2, 100 μg ml −1 BSA and 50 mM NaCl (NEB 2.1 buffer) at 37 °C for 2 h. Reactions were stopped and cleaned with HighPrep PCR cleanup beads (MAGBIO Genomics) and eluted in Tris-EDTA pH 8.0 buffer.

Article Title: Epistatic interaction between ribosome-associated Era GTPase and stringent response regulator RelA modulates bacterial cell growth and dormancy
Article Snippet: .. For in vitro translation assay (IVT), we used the PURExpress® Δ Ribosome Kit (NEB) and the reactions were set according to the prescribed protocol. ..

Article Title: LonP1 chaperone-like activity is ATPase independent and is mediated by its N-domain
Article Snippet: All plasmids were purchased from GeneScript. .. In vitro expression of the membrane protein OXA1L (72-435) and soluble protein DHFR was performed with the New England Biolabs PURExpress® In Vitro Protein Synthesis kit. .. The plasmid for DHFR was provided with kit, and the plasmid for OXA1L was purchased from GeneScript.

Expressing:

Article Title: Intrinsically disordered protein droplet-enhanced oligonucleotide assembly enables rapid oligonucleotide-to-protein expression
Article Snippet: .. Cell-free protein expression was performed using NEB PURExpress system supplemented with AlexaFluor647 (1.25 μM) and RNase inhibitor (2 U/μl). .. Reactions were encapsulated with fluorinated oil (AE3000 + 0.10% S386; AGC), sealed with Fomblin (Solvay) and incubated at 30°C for up to 16 h. Fluorescence imaging was performed using a Nikon TiE microscope (60× oil objective) with appropriate filter sets for AlexaFluor647, mNeonGreen, and mScarlet detection.

Membrane:

Concentration Assay:

Article Title: Detection of SARS-CoV-2 using RNA multi-arm junction logic gates
Article Snippet: .. Cell-Free Reactions Cell-free transcription-translation systems (NEB, PURExpress) were prepared for freeze-drying according to following recipe: cell-free solution A, 40%; cell-free solution B, 30%; RNase Inhibitor (Roche, 03335402001, distributed by MilliporeSigma), 2%; chlorophenol red-b-D-galactopyranoside (Roche, 10884308001, distributed by MilliporeSigma, 24 mg/ml), 2.5%; with the remaining volume reserved for LIRA riboregulator or gate RNA plasmids, water, and lacZ a peptide added to a final concentration of 2 μM. ..

Purification:

Article Title: Role of the anti-Shine–Dalgarno sequence of 16S rRNA in Flavobacterium johnsoniae
Article Snippet: .. A customized PURExpress ® (NEB) kit (lacking ribosomes and initiation factors) was supplemented with purified F. johnsoniae ribosomes and F. johnsoniae initiation factors and used to measure the synthesis of radiolabeled GFP fusion protein products, as described previously [ ]. .. A customized PURExpress (NEB) kit (lacking ribosomes and initiation factors) was supplemented with purified F. johnsoniae ribosomes and F. johnsoniae initiation factors and used to measure the synthesis of radiolabeled GFP fusion protein products, as described previously [ ].

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