pUC19 is a commonly used cloning vector that conveys the Amp resistance. The molecule is a small double-stranded circle, 2686 base pairs in length, and has a high copy number. pUC19 carries a 54 base-pair multiple cloning site polylinker that contains unique sites for 13 different hexanucleotide-specific restriction endonucleases (1) Plasmid pUC19 from Dr. Joachim Messing's lab is published in Gene. 1983 Dec;26(1):101-6. This plasmid is available through Addgene pUC18 / pUC19 from E.coli are commonly used small, high copy number, E. coli plasmids, Detaisl view of the pUC18/pUC19 product description. Reference 1. Yanisch-Perron, C., Vieira, J. and Messing, J. (1985) Gene 33, 103-119. 2. Accession L09137 X02514, Medline 85180545, Pubmed 2985470. Stability Very stable in the delivered form. The DNA is delivered frozen in storage buffer. Please. pUC19 Sequences (4) Addgene Sequences: Full (1) Partial (2) Depositing Scientist Sequences: Full (1) Full Sequences from Addgene (1) Based on next-generation sequencing (NGS) results where indicated (Addgene NGS Result), or assembled from reference sequences and/or Sanger results (Addgene Assembled Sequence). Analyze Sequence GenBank; SnapGene; File Help; Full Sequences from Depositor (1. pUC19 is one of a series of plasmid cloning vectors created by Joachim Messing and co-workers. The designation pUC is derived from the classical p prefix (denoting plasmid ) and the abbreviation for the University of California, where early work on the plasmid series had been conducted
.dna. Map and Sequence File: Download Open . Sequence Author: New England Biolabs. Download Free Trial Get SnapGene Viewer. Search. Your time is valuable! Basic Cloning Vectors. Plasmid Sets. Basic Cloning Vectors;; CRISPR Plasmids; Fluorescent Protein Genes & Plasmids; Gateway ® Cloning Vectors; I.M.A.G.E. Consortium Plasmids; Insect Cell Vectors; Luciferase Vectors; Lucigen. See Help:References for how to manage references in EcoliWiki. ↑ Yanisch-Perron, C et al. (1985) Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene 33 103-19 PubMed; Sequence 1 TCGCGCGTTT CGGTGATGAC GGTGAAAACC TCTGACACAT GCAGCTCCCG GAGACGGTCA CAGCTTGTCT GTAAGCGGAT GCCGGGAGCA GACAAGCCCG 101 TCAGGGCGCG TCAGCGGGTG TTGGCGGGTG.
The DNA molecular weight (MW) standard pUC19/ MspI allows the size comparison of gel-electrophoretical separated DNA-fragments in agarose or polyacrylamid gels. The reference range includes 12 defined DNA-bands with increasing base-pair length and is suited for equalizing of (in most cases unknown) DNA-fragments up to ca. 1000 bp. The MW. Reference: 1. Yanisch-Perron, C., Vieira, J. and Messing, J. (1985) Gene 33, 103-119. pUC19 Vector N3041S 50 µg Lot: 0371308 Exp: 8/15 1,000 µg/ml Store at -20°C Description: pUC19 is a commonly used plasmid cloning vector in E. coli. The molecule is a small double-stranded circle, 2686 base pairs in length, and has a high copy number. pUC19 carries a 54 base-pair multiple cloning site.
pUC19 est un plasmide développé par Yanisch-Perron, Vieira et Messing, à l'Université de Californie.Le nom du plasmide fait d'ailleurs référence à cette université (UC, pour University of California, p pour Plasmide). Ce plasmide, d'une longueur de 2686pb est un plasmide circulaire, à double brin .pUC19 est l'un des vecteurs de clonage les plus utilisés, dans la mesure où les. Cloning vector pUC19 Taxonomy ID: 31851 (for references in articles please use NCBI:txid31851) current name. Cloning vector pUC19. NCBI BLAST name: other sequences Rank: species Genetic code: Translation table 11 (Bacterial, Archaeal and Plant Plastid) Plastid genetic code: Translation table 11 (Bacterial, Archaeal and Plant Plastid) Other names: heterotypic synonym. Plasmid pUC19. Lineage. Plasmid DNA from E. coli RRI (pUC19, buffered aqueous solution); Plasmid DNA from Escherichia coli RRI has been used for imaging of DNA (nanostructure) via atomic force microscopy; The pUC19 plasmid (2,686 bp) confers ampicillin resistance and complement defects in β-galactosidase in ap
Reference: 1. Yanisch-Perron, C., Vieira, J. and Messing, J. (1985) Gene 33, 103-119. pUC19 Vector N3041S 50 µg Lot: 0371401 Exp: 1/16 1,000 µg/ml Store at -20°C Description: pUC19 is a commonly used plasmid cloning vector in E. coli. The molecule is a small double-stranded circle, 2686 base pairs in length, and has a high copy number. pUC19 carries a 54 base-pair multiple cloning site. Reference; Relationships; Motivational; Men's Health; Plasmid Cloning Vector pUC19. Last Updated on Mon, 07 Dec 2020 | Inherited Diseases. The plasmid pUC19, which is maintained in the bacterium E. coll, is a generalpurpose vector often used for cloning DNA fragments up to about 8kb in length. This vector is well conceived and useful for illustrating the basic principles of vector-based. dh5α/puc19 12 F-, Δ(argF-lac)169 , φ80d lacZ58 (M15), ΔphoA8 , glnX44 (AS) , λ - , deoR481 , rfbC1 ?, gyrA96 (NalR) , recA1 , endA1 , thiE1 , hsdR17 , pUC19
Location of important features in the pUC19 cloning vector plasmid. A p R is the ampicillin resistance gene, lacZα is the lacZ gene, and ori is the origin of replication. The numbers represent base pairs measured from an arbitrary reference point on the vector, labeled 0. The basepair numbers increase clockwise until the reference point is reached again after 2686 bp.. Strain DH5α/pUC19. DH5 is a non-mutagenized derivative of DH1, which transforms more efficiently due to a deoR mutation pUC18 and pUC19 vectors. The pUC18 and pUC19 plasmids enable successful cloning of large DNA fragments (larger than those cloned with a M13 mp18 RF Phage Vector). These cloning vectors contain a multiple cloning site at the lacZ' region that enables recombinant plamids to be verified via blue/white colony screening using agar plates containing IPTG. Reference: FGB 2001 Vector: puc19 Notes: for genomic library construction. Gene: niiA/AMA1 Name: pRG3-AMA1-niiA Size in kb: 11 Resistance: amp Depositor: GSM Reference: FGB 2001 Vector: puc19 Notes: for genomic library construction. Gene: pyr4 Plasmid name: AMA-NotI Size in kb:10 Kb Resistance:Amp Depositor: G.S. May Reference: Genetics 155:647-656 Vector: pUC19 The pUC19 was prepared by digestion with the EcoRI and HindIII restriction enzymes, resulting in a 2639 bp linear plasmid. Homologous recombination between the natMX and pUC19 fragments generated the pUC19Nat plasmid. (B) Agarose gel electrophoresis after gel purification of the fragments natMX and pUC19. (C) The counting of colonies after transformation of the vector alone and co-transformation of the pUC19 plus the fragment natMX. (D) Colony PCR screening confirmed 100% positive.
DH5α/pUC19 was included as a control to determine staining levels of live and heat-treated dead cells. Samples were excited with a 488 nm air-cooled argon ion laser in the CyFlow SL flow cytometer (Partec GmbH). Threshold settings were enabled on forward scatter to exclude cell debris. The forward and side scatter dot plot was used to identify and gate cell populations. Fluorescence was measured at 520 nm. Viable and dead cell populations were counted using the Partec, FloMax. Find SARS-CoV-2 related resources at NCBI. Download and submit sequences. Explore literature, identify clinical trials, and compounds used in them The plasmid must contain the lacZα, and examples of such plasmids are pUC19 and pBluescript. The E. coli cell should contain the mutant lacZ gene with deleted sequence (i.e. lacZΔM15), and some of the commonly used cells with such genotype are JM109, DH5α, and XL1-Blue. It should also be understood that the lac operon is affected by the presence of glucose. The protei pUC19 is one of a series of plasmid cloning vectors created by Joachim Messing and co-workers. The designation pUC is derived from the classical p prefix (denoting plasmid) and the abbreviation for the University of California, where early work on the plasmid series had been conducted.It is a circular double stranded DNA and has 2686 base pairs.. Transformation of non-competent Escherichia coli JM109 was accomplished using pUC19 as donor plasmid and sepiolite as the acicular material to promote cell piercing via application of friction with a polystyrene stick or a magnetic bar on the surface of a hydrogel containing agar. An automatic spreading setup was built with a conventional stirring plate and compared to manual spreading. Several parameters were optimized, namely, the agar content of the hydrogel (2%), concentration.
CHROM Name of the reference sequence. (CHROM is short for chromosome, a term that is not actually relevant here.) All results here used the pUC19 reference sequence. POS Position; the numbered location of the nucleotide in the sequence REF Reference nucleotide (A, C, T, or G) in the non-mutated reference sequence ALT Alternate nucleotide (A, C, T, or G) in the mutated sample sequence. If you wish to see recipes for making competent cells, consult the Sambrook reference in the lab. Each group will do 5 transformations: the test ligation and 4 controls. The controls will be the control ligation, unligated PstI-cut pUC19 vector DNA, pPBH plasmid miniprep (from Lab 3), and a mock transformation with no DNA added
These kanamycin resistant pUC vectors also contain a pUC19-derived multiple cloning site (MCS) within the lacZ gene, enabling recombinant clones to be verified through culture plates containing IPTG and X-Gal. High target gene expression is enabled by the presence of the lac promoter in both the pHSG298 and pHSG299 vectors EMBOSS Matcher identifies local similarities in two input sequences using a rigorous algorithm based on Bill Pearson's lalign application, version 2.0u4 (Feb. 1996 a, Time-course treatment of pUC19 DNA by SspE. In this assay, 0.3 µg of pUC19 DNA was untreated or treated with 2 µM SspE at 28 °C in CutSmart (New England Biolabs, 50 mM potassium acetate, 20. pg pUC19 DNA x µg x X µl plated x dilution factor For example, if transformation of 10 pg of pUC19 DNA yields 50 colonies when 25 µl of a 1:10 dilution is plated, then the transformation efficiency is: 50 colonies 106 pg 300 µl 10 pg DNA x µg x 25 µl plated x 10 = 6 x 108 Information for European Customers The ccdB Survival™ E. coli strain is genetically modified and carries the P trc.
The wild-type polymerase successfully amplified pUC19 DNA at standard 30°C temperature (up to 10 000-fold amplification after 6 hrs), meanwhile the MDA reaction carried at 40°C completely failed. In the same experiment the mutant enzyme (Mut) was able to perform MDA reaction both at 30 and 40°C. The accumulation of MDA product using mutant polymerase at 40°C was very steep during the first hours and reached the peak after 3 h (up to 30 000-fold amplification). The efficiency of MDA.
Regions of interest in the pUC19 plasmid Region Range LacZ alpha peptide 216-539 Lac operator (repressor binding site) 175-198 LacZ promoter 142-171 Ampicillin cassette 885-1745 Origin of replication 1900-2519 Mutation within 216-539 Example: No mutation within 216-53 A restriction endonuclease analysis of the bacterial plasmid controlling the EcoRI restriction and modification of DNA. Fed. Proc. 35, 2037-2043. PubMed Google Scholar. 23. Schickel, J., Helmig, C., and Meinhardt, F. (1996) Kluyveromyces lactis killer system: analysis of cytoplasmic promoters of the linear plasmids
Negative supercoiling in the 2686 bp Escherichia coli plasmid pUC19 is comparable in linking number (Lk0 = 258) and superhelical density (σ = −0.05) to the moderate supercoiling exhibited by many eukaryotic chromosomal DNAs in vivo. Supercoiled and relaxed forms of purified pUC19 in aqueous solution (0.1 M NaCl, pH 8.3, 20 °C) have been investigated by Raman spectroscopy to assess changes. Find protocols, product documentation, software, FAQs, videos, webinars, and more. Type in your search terms, and refine the results using the filters options on the left References: 1. Hanahan, D. (1983) J. Mol. Biol. 166, 557. 2. Tartof, K. D. and Hobbs, C. A. (1987) Focus ® 9 A stock pUC19 solution (0.01 µg/ml) is provided as a control to determine the transformation efficiency. The stock solution of pFastBac ™-gus (0.2 g/ml), µ provided with pFastBac™1 Expression Vector (Cat. No. 10360-014), can be used as a control for the transposition. The amplicons sets must include one end that overlaps by 20-22 bases with distinct ends of the pUC19 backbone. Import Plasmid mUAV (addgene) and Plasmid pUC19 (addgene) into Benchling. Benchling tutorials; Restriction digest pUC19 with PvuII and identify the backbone you want to use for your assembly. [Hint: You need a selection marker and origin of replication!] Identify the gene encoding for.
The Positive Control DNA Mix for Gibson Assembly consists of a two-piece assembly of pUC19. It is designed such that 5uL of the Positive Control DNA Mix is to be added to 15uL of Gibson Assembly Master Mix along side experimental reactions. Both pUC19 segments are between 1.3kb and 1.4kb in size. To construct the positive control reaction mix: PCR amplify the two pUC19 fragments - fragment 1. Sequence Extractor: Main | Features | Help | Download | License | About: Sequence Extractor generates a clickable restriction map and PCR primer map of a DNA sequence. Protein translations and intron/exon boundaries are also shown Reference management. Direct link. Watch-list. Remove from watch-list. Share this by email. Share this on Twitter. Share this on Facebook. Share this on Whatsapp . Export RIS Export BibTeX Export EndNote. Media type: Book; Thesis Title: Wirkungen der solaren UV-Strahlung auf das Escherichia-coli-Plasmid pUC19: Mutagenese, Inaktivierung und DNA-Strangbruchinduktion : 69 Tabellen Contributor.
10pg / μL solution of a standard plasmid (e.g. pUC19) SOC Medium; PROTOCOL: ) Transform 10pg of a standard plasmid (e.g. pUC19) in 50μL of cells using the standard transformation protocol. ) Plate 50uL of transformed cells in triplicate on appropriate antibiotic ) Grow plates overnight, and count colonies the next morning ) Average colony counts for the three plates; efficiency = (# colonies. pUC19 Permits and Restrictions: View Permits. Depositors J Messing References Vieira J, Messing J. The pUC plasmids, an M13mp7-derived system for insertion mutagenesis and sequencing with synthetic universal primers. Gene 19: 259-268, 1982. PubMed: 6295879. Norrander J, et al. Construction of improved M13 vectors using oligodeoxynucleotide-directed mutagenesis. Gene 26: 101-106, 1983. (B) CBS7435 harboring pUC19-Cen2-EGFP (Cen2), pUC19-LOR2CC2-EGFP (LOR2-CC2), pUC19-LOR2-EGFP (LOR2), pUC19-LOR2(1-111)-EGFP [LOR2(1-111)], and pUC19-PARS1-EGFP (PARS1) was grown in BMGY medium with or without 200 μg/ml Zeocin for 24 h. The strains CBS7435 (control) and T38473-EGFP (genomic integration) were grown in BMGY medium without Zeocin for 24 h. The GFP expression level of each. Abstract. The development of gel electrophoresis as a method of separating and analyzing DNA has been one of the forces driving the revolution in molecular biology for the last 20 years. In principle, DNA gel electrophoresis is conceptually easy to understand and technically easy to execute. In practice, there are a lot of small details that affect.
We performed a screen for extrachromosomal circular DNAs containing segments of genomic yeast DNA. We found 1,756 such extrachromosomal circular DNAs containing about 23% of the total yeast genomic information. The abundance of these circular forms of genomic DNA suggests that eccDNA formation might be a common mutation that can arise in any part of the genome, and not in only a few special loci Cloning vector pUC19-35S-FLAG-RBS: Taxonomy navigation › vectors. Terminal (leaf) node. Common name i-Synonym i-Rank i: SPECIES : Lineage i › other. Molekularbiologische Untersuchungen der am E. coli Plasmid pUC19 durch UVA und UVB induzierten Strahlenschaeden ASCII Citation Atom BibTeX Dublin Core EP3 XML EndNote Grid (abstract) HTML Citation JSON METS MODS MPEG-21 DIDL Multiline CSV OpenURL ContextObject OpenURL ContextObject in Span RDF+N-Triples RDF+N3 RDF+XML Refer Reference Manager Simple Metadat pUC19 Control DNA for NEB 5-alpha Competent E. coli Required Materials Not Included: DNA Polymerase (for generating PCR products): We recommend Q5® High-Fidelity DNA Polymerase (NEB #M0491) or related products, such as Q5 Hot Start High-Fidelity DNA Polymerase (NEB #M0493) or Q5 Hot Start High-Fidelity 2X Master Mix (NEB #M0494)
Reference Sequence. Ordering: Name Cat. No. Availability; pUC19-HRP-C: MC_0003747: Digital Sequence: please Send Request for availability : Send Request Edit in GenSmart Design: This material may be covered by one or more patents, trademarks and/or copy rights owned or controlled by Depositors or any third parties. This material is available to academic and nonprofit organizations for. pBK-CMV Phagemid Vector 1 pBK-CMV Phagemid Vector MATERIALS PROVIDED Material provided Quantity Storage pBK-CMVa phagemid vector(1 μg/μl) 20 μg -20°C XL1-Blue MRF´b 500 μl -80°C R408 Interference-Resistant Helper Phage 1 ml -80° We digested the pUC19 vector by BamHI and EcoRI and purified the digested vector on gel. We proceeded to the InFusion kit reaction, transformation of Stellar cells, selection on ampicillin, and minipreps from 6 clones. The plasmids were assessed by restriction profiling with the enzymes BamHI and EcoRI. Figure 7: Verification using the InFusion Kit: the expected sizes were 4.8 kb and 2.6 kb. JM109 is a K strain that is recA- and endA- to minimize recombination and improve the quality of plasmid DNA.In addition, the cells carry the F´ episome, which allows blue/white screening. JM109 Competent Cells are available for convenient transformation in two efficiencies: High Efficiency at greater than 10 8 cfu/μg for Cloning and Single Use, and Subcloning Efficiency at greater than. Reference: Yanisch-Perron, C, J Vieira, J Messing 1985. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors
pUC19 is a small, high-copy number E. coli plasmid cloning vector containing portions of pBR322 and M13mp19 (1). It con-tains the pMB1 origin of replication from pBR322, but it lacks the rop gene and carries a point mutation in the RNAII transcript (G 2975 in pBR322 to A 1308 in pUC19; 2). These changes together result in a temperature-dependent copy number of about 75 per cell at 37°C and. PUC19 2015-01-13 azim58 - PUC19 PUC19 plasmid map: http://www.neb.com/nebecomm/tech_reference/restriction_enzymes/maps/puc19_m ap.pdf 2686 bp encodes for beta. Briefly, primers pUC19_F (5′‐gcatga AAGCTT GCATGCCTGCAGGTCGAC‐3′) and pUC19_R (5′‐gcatga CATATG CGGTGTGAAATACCGCAC‐3′), which incorporate a HindIII and NdeI site, respectively (underlined), were used to amplify a 265 bp fragment of the lacZα gene using pUC19 as a template This quick reference sheet is provided for experienced users of the Gateway pUC19 Control DNA 10 pg/ μL in 5 mM Tris-HCl, 0.5 mM EDTA, pH 8. 50 μL . Genotype of OmniMAX™ 2-T1R. F′ [proAB+ lacIq lacZΔM15 Tn10(TetR) Δ(ccdAB)] mcrA mrr-hsdRMS- BC) φ80(lacZ)ΔM15 Δ(lacZYA-argF) U169 endA1 recA1 supE44 thi-1 gyrA96 relA1 tonA panD . Introduction . Overview . Introduction . The Gateway.
pUC19 is a plasmid cloning vector created by Messing and co-workers in the University of California. p in the name stands for plasmid and UC represents the University in which it was created.It is a circular double stranded DNA and has 2686 base pairs . pUC19 is one of the most widely used vector molecules as the recombinants, or the cells into which foreign DNA has been introduced, can be. In addition, pUC19 plasmid was extracted from the transformed strain, and the AS-1 wild-type strain became resistant to ampicillin by reintroduction of this plasmid. Thus, the AS-1 strain was able to express the ampicillin resistance gene encoded by the pUC19 plasmid. Which led us to conclude that strain AS-1 could be used as a eurytrophic recombinant host Verify the sequence of each colony by sequencing from the pUC19 backbone using the pUC19-Fwd or pUC19-Rev primer. Reference the sequencing results against the expected genomic sequence to check. privacy notice: disclaimer: accessibility: application suppor
model.GATC.R9_2D.tem.puc19.bn17.sn360.tar.gz: A GATC model trained using pUC19 R9 2D template reads (for deepsignal<=0.1.6). Example data. The example data can be downloaded from google drive. fast5s.sample.tar.gz: The data contain ~4000 yeast R9.4 1D reads each with called events (basecalled by Albacore), along with a genome reference. Quick star user guide pcDNA™3.4-TOPO® TA Cloning® Kit Five-minute cloning and expression of Taq polymerase-amplified PCr products in mammalian cells Catalog Number A14697 Publication Number MAN0007209 Revision 2.
Reference Sequence. Ordering: Name Cat. No. Availability; pUC19/human IL-8: MC_0006459: Digital Sequence: please Send Request for availability : Send Request Edit in GenSmart Design: This material may be covered by one or more patents, trademarks and/or copy rights owned or controlled by Depositors or any third parties. This material is available to academic and nonprofit organizations. The unpolished control DNA (pUC19) contains 3´-end nucleotide extensions and will ligate at an extremely reduced efficiency in the presence of T4 DNA ligase. PCR polishing removes 3´-end nucleotide extensions created by DNA polymerases and will produce blunt-ended DNA molecule Unless otherwise cited or referenced, all content of this presenataion is licensed under the Creative Commons License Attribution Share-Alike 2.5 Canada. 2 Plasmids Naturally occurring plasmids-occur widely in bacteria -are covalently closed circular dsDNA-are replicons, stably inherited as extra-chromosomal DNA-can be 1 kbp to 500 kbp in size (compared to 4000 kbp chromosome)-bacteria can. References (45) Figures (5) (SC) pUC19 DNA into nicked circular (NC) and linear (LC) conformation can be used to quantify the relative cleavage efficiency of complexes by . agarose gel.
pUC19 plasmid DNA Description pUC19 is a common used plasmid cloning vector in E. coli. The vector length is 2686 bp and is isolated from E. coli strain DH5 a by standard procedures. Storage Buffer and Concentration Supplied as 500 ng/ml in TE buffer. Store at -20 °C. Polylinker Start: 0: Polylinker End: 0: u6 Start: 0: U6 End: 0: H1 Start: 0. There are two primary methods for transforming bacterial cells: heat shock and electroporation. In both cases, the bacterial cells have to be made competent or permeable to plasmids that you would like the cell to propagate . CRISPR/Cas13a (known previously as C2c2) is a class 2 type VI-A ribonuclease capable of targeting and cleaving single-stranded RNA (ssRNA) molecules of the phage genome. Here, we employ CRISPR/Cas13a to engineer interference with an RNA virus, Turnip Mosaic Virus (TuMV), in plants Salmonella enterica subsp. enterica serotype Enteritidis BM4361 and BM4362 were isolated from the same patient. BM4361 was susceptible to aminoglycosides, whereas BM4362 was resistant to tobramycin owing to synthesis of a 6′- N -acetyltransferase type I [AAC(6′)-I]. Comparative analysis of nucleotide sequences, pulsed-field gel electrophoresis patterns, and Southern hybridizations.
Information on EC 220.127.116.11 - lysozyme. crystallization data and molecular dynamics simulations indicate that lysozyme is an inverting enzyme, and Asp97 acts as a second carboxylate and that the narrow space of the binding cleft at subsites E-G in GEL may prohibit the sugar chain to bind alternative site that might be essential for transglycosylatio References. 1. Richardson, Christopher D., et al. Enhancing homology-directed genome editing by catalytically active and inactive CRISPR-Cas9 using asymmetric donor DNA. Nature biotechnology (2016). PubMed PMID: 26789497. 2. Lin, Steven, et al. Enhanced homology-directed human genome engineering by controlled timing of CRISPR/Cas9 delivery. Elife 3 (2015): e04766. PubMed PMID: 25497837.
To find out the structure of any allele connecting the introduced sequences to the bovine chromosomes, a new reference was generated to include the bovine reference ARS-UCD1.2 , together with the pUC19 plasmid and complete template sequences. All candidate reads were aligned against the new reference. To enable better delineation of the allele structure, each read was fragmented into 1 kb. The resulting full-length products were amplified by PCR and cloned into the pUC19 vector. The ampicillin gene (amp) The comparative C q method was used to calculate the expression level of the target region relative to a reference region (ACTB locus). Percent of deletion in the target region was further calculated by the ratio of target cells relative to control cells. All primer. Take 5μL Puc19-recaegfp (hereinafter referred to as pre plasmid), the production of enzyme ligation, and transfer it into E.coli Trans5α and E.coli BL21 (DE3) competent cells under the same conditions above. Apply 100μL on LB plate which contains 100 μg/mL of ampicillin resistance, culture it upside down at 37℃ overnight, select the monoclonal colony, expand the culture, send it to. Plan du cours 4/183 Biologie génique - Pr A. Raisonnier 2006 - 2007 41 Chapitre 5 : Les nucléases 42 5.1 Fragment de restriction (définition) 43 5.2 Système de restriction-modification 44 5.3 Nomenclature des enzymes de restriction 45 5.4 Restriction (réaction générale) 46 5.5 Tampons d'incubation (restriction) 47 5.6 EcoR I 48 5.7 EcoR V 49 5.8 Pvu II 50 5.9 Hae II