Supporting Information. Copyright Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim, 2006
|
|
- Caitlin Gaines
- 6 years ago
- Views:
Transcription
1 Supporting Information Copyright Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim, 2006
2 Copyright Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim, 2006 Supporting Information for Expanding the Genetic Code in a Mammalian Cell by the Introduction of 4-Base Codon/Anticodon Pairs Masumi Taki, Jiro Matsushita and Masahiko Sisido* Experimental Section 1. Construction of plasmids that express EGFPs 1.1. Plasmid for wild-type EGFP: An NheI restriction site, a Kozak sequence for translation initiation with an ATG start codon, and a FLAG-tag were inserted by PCR just before the initiation codon of the EGFP-encoding gene that originated from pegfp-n3 (Clontech). A HindIII restriction site was also added just after the stop codon of the gene. By using the NheI and HindIII sites, the amplified gene was subcloned into a vector (pcdna5/frt; Invitrogen, Carlsbad, CA) to generate a plasmid that expresses wild-type EGFP (pcdna5/ FRT/EGFPwt) Plasmids for amber mutated EGFP: First, the 66 TAC codon of wild-type EGFP was replaced by an amber codon "TAG" using an improved site-directed mutagenesis based on a QuikChange TM protocol. The sequences of the sense and antisense primers were, 5'- GAC CAC CCT GAC CTA GGG CGT GCA GTG C-3' and 5'-GCA CTG CAC GCC CTA GGT CAG GGT GGT C-3', respectively, with the mutated positions being under-lined. Note 1
3 that site-directed mutagenesis was successfully carried out simply by using the primer pair whose melting temperature on both sides of the mutation position is estimated to be C, from the contents of G:C (+4 C) and A:T (+2 C) pairs [B. Cormack, Current Protocols in Molecular Biology, Wiley, New York, 1991, ]. These primers were purchased from Hokkaido System Science Co., Ltd. (Sapporo, Japan). The site-directed mutation was successful with the primers that were not puri-fied by HPLC, but purified simply and inexpensively by using a reverse-phase cartridge. The 50 µl-scale PCR was carried out with 250 ng of the template DNA (pcdna5/frt/egfpwt), 0.3 µm of the primer pair, 500 µm of dntps and 2.5 U of Pfu Turbo DNA polymerase (Stratagene). The PCR was initiated by preheating the reaction mixture at 95 C for 1 min, followed by 16 cycles of 94 C for 30 s, 55 C for 1 min and 68 C for 12 min. The resulting PCR mixture was treated with a restriction enzyme DpnI (New England Biolabs) for 1 h without further purification in the same PCR tube, and an aliquot (1 µl) of the mixture was directly transformed into DH5α chemocompetent cells. The transformed cells were cultivated on a Luria-Bertani (LB) plate containing ampicillin. The colonies were selected and their plasmids were isolated by a Plasmid Miniprep Kit (Bio-Rad), to obtain the mutated plasmid (pcdna5/frt/ EGFP 66TAG ) Plasmids for EGFPs that contain 66 UAGN, 66 CNCU, 66 CUAU and 66 UUGU 4-base codons, and that contain 66 UAGNN 5-base codons: The plasmid for the EGFP that contain 66 UAGN 4-base codons was prepared by essentially the same procedure as described above, except for using pcdna5/frt/egfp 66TAG as the template plasmid and using the following mixed oligonucleotides as sense primers, 5'-GAC CAC CCT GAC CTA GNG GCG TGC AGT GC-3'; and as antisense primers, 5'-GCA CTG CAC GCC NCT AGG TCA GGG TGG TC-3'. A total of 12 colonies were selected, and their plasmids were isolated by a Plasmid Miniprep Kit and sequenced. All four mutants that con-tain one of the TAGN codons were obtained through this one-step transformation, i.e., four independent EGFP-expression vectors containing the four different types of 4-base codons (pcdna5/ FRT/EGFP 66TAGN ) were simultaneously generated in a single PCR tube from a single template vector that contains an amber codon (pcdna5/frt/egfp 66TAG ). The plasmids for the EGFP that contain 66 CNCU, 66 CUAU, 66 UUGU, and 66 UAGNN codons, were also prepared by essentially the same procedure as above, except for using the following oligonucleotides: 5'-GAC CAC CCT GAC CCN CTG GCG TGC AGT GC-3' 2
4 and 5'-GCA CTG CAC GCC AGN GGG TCA GGG TGG TC-3' for 66 CNCU; 5'-GAC CAC CCT GAC CCT ATG GCG TGC AGT GC-3' and 5'-GCA CTG CAC GCC ATA GGG TCA GGG TGG TC-3' for 66 CUAU; 5'-GAC CAC CCT GAC CTT GTG GCG TGC AGT GC-3' and 5'-GCA CTG CAC GCC ACA AGG TCA GGG TGG TC-3' for 66 UUGU; and 5'-CAC CCT GAC CTA GNN GGC GTG CAG TGC-3' and 5'-GCA CTG CAC GCC NNC TAG GTC AGG GTG GTC-3' for 66 UAGNN. 2. Construction of trna-expressing plasmids 2.1. Plasmid for trna that contains an amber anticodon: A pgem-t-tr-hutyr(cta) plasmid was designed to encode 5 - and 3 -flanking sequences derived from the human trna Tyr gene and a trna-expressing sequence containing an internal promoter and an amber anticodon. The following oligonucleotides were annealed: forward, 5'-GGT TTT TCT GTG CTG AAC CTC AGG GGA CGC CGA CAC ACG TAC ACG TCC CTT CGA TAG CTC AGC TGG TAG AGC-3'; reverse, 5'-GCA CTT GTC TCC TTC GAG CCG GAA TCG AAC CAG CGA CCT AAG GAT CTA GAG TCC TCC GCT CTA CCA GCT GAG C-3'. The primer extension reaction was performed by using a thermal cycler with Ex Taq TM DNA polymerase (Takara, Shiga, Japan). By using this reaction product as the template DNA, PCR was carried out with a sense primer, 5'-GGC TCT AGA ACC TGC CGG CCA CCA GCG CTC CGG TTT TTC TGT GCT GAA CC' and an antisense primer, 5'-CTG CCC GAC CTG CTC ACG CAT GAA AAA AAC CGC ACT TGT CTC CTT CGA G-3'. The PCR product was cloned into pgem-t Easy vector (Promega) according to the manufacturer s protocol to obtain pgem- T-tR-HuTyr(cta) Plasmids for trnas that contain 4-base anticodons (ncua, agng, auag, and acaa), and 5-base anticodons (cccua): The plasmids for trnas that contain one of the 4-base anticodons, ncua, were prepared by essentially the same procedure as described above, except for using pgem-t-tr-hutyr(cta) as a template plasmid and the following oligonucleotides as a sense primer, 5'-GAC CTA AGG ATC TAG NAG TCC TCC GCT CTA C- 3'; and as an antisense primer, 5'-GTA GAG CGG AGG ACT NCT AGA TCC TTA GGT C- 3'. The colonies were selected, their plasmids were isolated by Plasmid Miniprep Kit and sequenced. All four mutants of pgem-t-tr-hutyr(ncta) that contained ncta anticodons were obtained through this one-step transformation. The plasmids for trnas that contain 4-base anticodons agng, auag, acaa, and 5-base 3
5 anticodon cccua were prepared by essentially the same procedure as described above, except for using the following oligonucleotides, 5'-GCG ACC TAA GGA TCC NCT AGT CCT CCG CTC TA-3' and 5'-TAG AGC GGA GGA CTA GNG GAT CCT TAG GTC GC-3' for agng; 5'-GCG ACC TAA GGA TCC TAT AGT CCT CCG CTC TAC-3' and 5'-GTA GAG CGG AGG ACT ATA GGA TCC TTA GGT CGC-3' for auag; 5'-GCG ACC TAA GGA TCT TGT AGT CCT CCG CTC TAC-3' and 5'-GTA GAG CGG AGG ACT ACA AGA TCC TTA GGT CGC-3' for acaa; 5'-GAC CTA AGG ATC TAG GGA GTC CTC CGC TCT-3' and 5'- AGA GCG GAG GAC TCC CTA GAT CCT TAG GTC-3' for cccua. 4
Electronic Supplementary Information
Electronic Supplementary Material (ESI) for Molecular BioSystems. This journal is The Royal Society of Chemistry 2017 Electronic Supplementary Information Dissecting binding of a β-barrel outer membrane
More informationSupplement 1: Sequences of Capture Probes. Capture probes were /5AmMC6/CTG TAG GTG CGG GTG GAC GTA GTC
Supplementary Appendixes Supplement 1: Sequences of Capture Probes. Capture probes were /5AmMC6/CTG TAG GTG CGG GTG GAC GTA GTC ACG TAG CTC CGG CTG GA-3 for vimentin, /5AmMC6/TCC CTC GCG CGT GGC TTC CGC
More informationFigure S1. Characterization of the irx9l-1 mutant. (A) Diagram of the Arabidopsis IRX9L gene drawn based on information from TAIR (the Arabidopsis
1 2 3 4 5 6 7 8 9 10 11 12 Figure S1. Characterization of the irx9l-1 mutant. (A) Diagram of the Arabidopsis IRX9L gene drawn based on information from TAIR (the Arabidopsis Information Research). Exons
More informationTable S1. Bacterial strains (Related to Results and Experimental Procedures)
Table S1. Bacterial strains (Related to Results and Experimental Procedures) Strain number Relevant genotype Source or reference 1045 AB1157 Graham Walker (Donnelly and Walker, 1989) 2458 3084 (MG1655)
More informationSupplemental Data. mir156-regulated SPL Transcription. Factors Define an Endogenous Flowering. Pathway in Arabidopsis thaliana
Cell, Volume 138 Supplemental Data mir156-regulated SPL Transcription Factors Define an Endogenous Flowering Pathway in Arabidopsis thaliana Jia-Wei Wang, Benjamin Czech, and Detlef Weigel Table S1. Interaction
More informationAdd 5µl of 3N NaOH to DNA sample (final concentration 0.3N NaOH).
Bisulfite Treatment of DNA Dilute DNA sample to 2µg DNA in 50µl ddh 2 O. Add 5µl of 3N NaOH to DNA sample (final concentration 0.3N NaOH). Incubate in a 37ºC water bath for 30 minutes. To 55µl samples
More informationstrain devoid of the aox1 gene [1]. Thus, the identification of AOX1 in the intracellular
Additional file 2 Identification of AOX1 in P. pastoris GS115 with a Mut s phenotype Results and Discussion The HBsAg producing strain was originally identified as a Mut s (methanol utilization slow) strain
More informationSupplementary Materials for
www.sciencesignaling.org/cgi/content/full/10/494/eaan6284/dc1 Supplementary Materials for Activation of master virulence regulator PhoP in acidic ph requires the Salmonella-specific protein UgtL Jeongjoon
More informationSAY IT WITH DNA: Protein Synthesis Activity by Larry Flammer
TEACHER S GUIDE SAY IT WITH DNA: Protein Synthesis Activity by Larry Flammer SYNOPSIS This activity uses the metaphor of decoding a secret message for the Protein Synthesis process. Students teach themselves
More informationSupplementary Information. Construction of Lasso Peptide Fusion Proteins
Supplementary Information Construction of Lasso Peptide Fusion Proteins Chuhan Zong 1, Mikhail O. Maksimov 2, A. James Link 2,3 * Departments of 1 Chemistry, 2 Chemical and Biological Engineering, and
More informationII 0.95 DM2 (RPP1) DM3 (At3g61540) b
Table S2. F 2 Segregation Ratios at 16 C, Related to Figure 2 Cross n c Phenotype Model e 2 Locus A Locus B Normal F 1 -like Enhanced d Uk-1/Uk-3 149 64 36 49 DM2 (RPP1) DM1 (SSI4) a Bla-1/Hh-0 F 3 111
More informationLecture 11: Gene Prediction
Lecture 11: Gene Prediction Study Chapter 6.11-6.14 1 Gene: A sequence of nucleotides coding for protein Gene Prediction Problem: Determine the beginning and end positions of genes in a genome Where are
More informationSupplemental Information. Target-Mediated Protection of Endogenous. MicroRNAs in C. elegans. Inventory of Supplementary Information
Developmental Cell, Volume 20 Supplemental Information Target-Mediated Protection of Endogenous MicroRNAs in C. elegans Saibal Chatterjee, Monika Fasler, Ingo Büssing, and Helge Großhans Inventory of Supplementary
More informationG+C content. 1 Introduction. 2 Chromosomes Topology & Counts. 3 Genome size. 4 Replichores and gene orientation. 5 Chirochores.
1 Introduction 2 Chromosomes Topology & Counts 3 Genome size 4 Replichores and gene orientation 5 Chirochores 6 7 Codon usage 121 marc.bailly-bechet@univ-lyon1.fr Bacterial genome structures Introduction
More informationCodon Bias with PRISM. 2IM24/25, Fall 2007
Codon Bias with PRISM 2IM24/25, Fall 2007 from RNA to protein mrna vs. trna aminoacid trna anticodon mrna codon codon-anticodon matching Watson-Crick base pairing A U and C G binding first two nucleotide
More informationSearch for and Analysis of Single Nucleotide Polymorphisms (SNPs) in Rice (Oryza sativa, Oryza rufipogon) and Establishment of SNP Markers
DNA Research 9, 163 171 (2002) Search for and Analysis of Single Nucleotide Polymorphisms (SNPs) in Rice (Oryza sativa, Oryza rufipogon) and Establishment of SNP Markers Shinobu Nasu, Junko Suzuki, Rieko
More informationSupplementary Information
Supplementary Information A general solution for opening double-stranded DNA for isothermal amplification Gangyi Chen, Juan Dong, Yi Yuan, Na Li, Xin Huang, Xin Cui* and Zhuo Tang* Supplementary Materials
More informationEngineering Escherichia coli for production of functionalized terpenoids using plant P450s
Supplementary Information for Engineering Escherichia coli for production of functionalized terpenoids using plant P450s Michelle C. Y. Chang, Rachel A. Eachus, William Trieu, Dae-Kyun Ro, and Jay D. Keasling*
More informationPROTEIN SYNTHESIS Study Guide
PART A. Read the following: PROTEIN SYNTHESIS Study Guide Protein synthesis is the process used by the body to make proteins. The first step of protein synthesis is called Transcription. It occurs in the
More informationEngineering D66N mutant using quick change site directed mutagenesis. Harkewal Singh 09/01/2010
Engineering D66N mutant using quick change site directed mutagenesis Harkewal Singh 09/01/2010 1 1- What is quick change site directed mutagenesis? 2- An overview of the kit contents. 3- A brief information
More informationSupplementary Material and Methods
Supplementary Material and Methods Synaptosomes preparation and RT-PCR analysis. Synaptoneurosome fractions were prepared as previously described in 1. Briefly, rat total brain was homogenized in ice-cold
More informationChapter 13 Chromatin Structure and its Effects on Transcription
Chapter 13 Chromatin Structure and its Effects on Transcription Students must be positive that they understand standard PCR. There is a resource on the web for this purpose. Warn them before this class.
More informationSupplemental Data. Lin28 Mediates the Terminal Uridylation. of let-7 Precursor MicroRNA. Molecular Cell, Volume 32
Molecular Cell, Volume 32 Supplemental Data Lin28 Mediates the Terminal Uridylation of let-7 Precursor MicroRNA Inha Heo, Chirlmin Joo, Jun Cho, Minju Ha, Jinju Han, and V. Narry Kim Figure S1. Endogenous
More informationSupplemental Data. Distinct Pathways for snorna and mrna Termination
Molecular Cell, Volume 24 Supplemental Data Distinct Pathways for snorna and mrna Termination Minkyu Kim, Lidia Vasiljeva, Oliver J. Rando, Alexander Zhelkovsky, Claire Moore, and Stephen Buratowski A
More informationSupporting Information. Trifluoroacetophenone-Linked Nucleotides and DNA for Studying of DNA-protein Interactions by 19 F NMR Spectroscopy
Supporting Information Trifluoroacetophenone-Linked Nucleotides and DNA for Studying of DNA-protein Interactions by 19 F NMR Spectroscopy Agata Olszewska, Radek Pohl and Michal Hocek # * Institute of Organic
More informationSupporting Information
Supporting Information Wiley-VCH 2007 69451 Weinheim, Germany Site-Specific Control of Distances between Gold Nanoparticles using Phosphorothioate Anchors on DNA and a Short Bifunctional Molecular Fastener
More informationA high efficient electrochemiluminescence resonance energy. transfer system in one nanostructure: its application for
Supporting Information for A high efficient electrochemiluminescence resonance energy transfer system in one nanostructure: its application for ultrasensitive detection of microrna in cancer cells Zhaoyang
More informationLezione 10. Bioinformatica. Mauro Ceccanti e Alberto Paoluzzi
Lezione 10 Bioinformatica Mauro Ceccanti e Alberto Paoluzzi Dip. Informatica e Automazione Università Roma Tre Dip. Medicina Clinica Università La Sapienza Lezione 10: Sintesi proteica Synthesis of proteins
More informationIn-Fusion Advantage PCR Cloning Kit
User Manual In-Fusion Advantage PCR Cloning Kit User Manual United States/Canada 800.662.2566 Asia Pacific +1.650.919.7300 Europe +33.(0)1.3904.6880 Japan +81.(0)77.543.6116 Clontech Laboratories, Inc.
More informationWet Lab Tutorial: Genelet Circuits
Wet Lab Tutorial: Genelet Circuits DNA 17 This tutorial will introduce the in vitro transcriptional circuits toolkit. The tutorial will focus on the design, synthesis, and experimental testing of a synthetic
More informationPILRα Is a Herpes Simplex Virus-1 Entry Coreceptor That Associates with Glycoprotein B
Satoh et al. Page S1 Cell, Volume 132 PILRα Is a Herpes Simplex Virus-1 Entry Coreceptor That Associates with Glycoprotein B Takeshi Satoh, Jun Arii, Tadahiro Suenaga, Jing Wang, Amane Kogure, Junji Uehori,
More informationSequence Design for DNA Computing
Sequence Design for DNA Computing 2004. 10. 16 Advanced AI Soo-Yong Shin and Byoung-Tak Zhang Biointelligence Laboratory DNA Hydrogen bonds Hybridization Watson-Crick Complement A single-stranded DNA molecule
More informationGenomics and Gene Recognition Genes and Blue Genes
Genomics and Gene Recognition Genes and Blue Genes November 1, 2004 Prokaryotic Gene Structure prokaryotes are simplest free-living organisms studying prokaryotes can give us a sense what is the minimum
More informationSUPPLEMENTARY INFORMATION. doi: /nature08559
Supplementary Materials and methods Genetic constructs: Construction of prophoa-his, prophoa(l8q)-his, prophoa(l14r)-his, PhoA(Δ2-22)-His and prophoa(1-62)-his for purification purposes: prophoa-his 6
More informationSupplemental Data. Short Article. Transcriptional Regulation of Adipogenesis by KLF4. Kıvanç Birsoy, Zhu Chen, and Jeffrey Friedman
Cell Metabolism, Volume 7 Supplemental Data Short Article Transcriptional Regulation of Adipogenesis by KLF4 Kıvanç Birsoy, Zhu Chen, and Jeffrey Friedman Supplemental Experimental Procedures Plasmids
More informationNucleic Acids Research
Volume 10 Number 1 1982 VoLume 10 Number 11982 Nucleic Acids Research Nucleic Acids Research A convenient and adaptable package of DNA sequence analysis programs for microcomputers James Pustell and Fotis
More informationCodon bias and gene expression of mitochondrial ND2 gene in chordates
www.bioinformation.net Hypothesis Volume 11(8) Codon bias and gene expression of mitochondrial ND2 gene in chordates Arif Uddin, Tarikul Huda Mazumder, Monisha Nath Choudhury & Supriyo Chakraborty* Department
More informationDNA extraction. ITS amplication and sequencing. ACT, CAL, COI, TEF or TUB2 required for species level. ITS identifies to species level
DNA extraction ITS amplication and sequencing ITS identifies to species level Ceratocystis fagacearum Ceratocystis virescens Melampsora medusae (only to species) Mycosphaerella laricis-leptolepidis Phaeoramularia
More informationInt J Clin Exp Med 2014;7(9): /ISSN: /IJCEM Jin Ah Ryuk *, Young Seon Kim *, Hye Won Lee, Byoung Seob Ko
Int J Clin Exp Med 2014;7(9):2488-2496 www.ijcem.com /ISSN:1940-5901/IJCEM0001438 Original Article Identification of Acorus gramineus, A. calamus, and A. tatarinowii using sequence characterized amplified
More informationINTRODUCTION TO THE MOLECULAR GENETICS OF THE COLOR MUTATIONS IN ROCK POCKET MICE
The Making of the The Fittest: Making of the Fittest Natural Selection Natural and Adaptation Selection and Adaptation Educator Materials TEACHER MATERIALS INTRODUCTION TO THE MOLECULAR GENETICS OF THE
More informationMOLECULAR CLONING AND SEQUENCING OF FISH MPR 46
MOLECULAR CLONING AND SEQUENCING OF FISH MPR 46 INTRODUCTION The mammalian MPR 46 (human, mouse, bovine) sequences show extensive homologies. Among the non-mammalian vertebrates, only a partial sequence
More informationMeixia Li, Chao Cai, Juan Chen, Changwei Cheng, Guofu Cheng, Xueying Hu and Cuiping Liu
S1 of S6 Supplementary Materials: Inducible Expression of both ermb and ermt Conferred High Macrolide Resistance in Streptococcus gallolyticus subsp. pasteurianus Isolates in China Meixia Li, Chao Cai,
More information2.5. Equipment and materials supplied by user Template preparation by cloning into plexsy_invitro-2 vector 4. 6.
Manual 15 Reactions LEXSY in vitro Translation Cell-free protein expression kit based on Leishmania tarentolae contains the vector plexsy_invitro-2 Cat. No. EGE-2002-15 FOR RESEARCH USE ONLY. NOT INTENDED
More informationElectronic Supplementary Information
Electronic Supplementary Information Ultrasensitive and Selective DNA Detection Based on Nicking Endonuclease Assisted Signal Amplification and Its Application in Cancer Cells Detection Sai Bi, Jilei Zhang,
More informationHigh-throughput cloning and expression in recalcitrant bacteria
High-throughput cloning and expression in recalcitrant bacteria Eric R Geertsma & Bert Poolman Supplementary text and figures: Supplementary Figure 1 Frequency of SfiI sites yielding identical 3 extensions
More informationCloning and characterization of a cdna encoding phytoene synthase (PSY) in tea
African Journal of Biotechnology Vol. 7 (20), pp. 3577-3581, 20 October, 2008 Available online at http://www.academicjournals.org/ajb ISSN 1684 5315 2008 Academic Journals Full Length Research Paper Cloning
More informationSupplemental Data. Sheerin et al. (2015). Plant Cell /tpc h FR lifetime (ns)
A CFP YFP Merge phya-cfp YFP-SPA1 D phya-cfp YFP-SPA1 6 h FR phya-nls-cfp YFP-SPA3 phya-nls-cfp YFP-SPA4 B n P value phya-nls-cfp 31 - phya-nls-cfp YFP-SPA3 8 0.02 phya-nls-cfp YFP-SPA4 9 0.48 1.6 1.8
More informationBest practices for Variant Calling with Pacific Biosciences data
Best practices for Variant Calling with Pacific Biosciences data Mauricio Carneiro, Ph.D. Mark DePristo, Ph.D. Genome Sequence and Analysis Medical and Population Genetics carneiro@broadinstitute.org 1
More informationAmplified Analysis of DNA by the Autonomous Assembly of Polymers Consisting of DNAzyme Wires
Supporting information for the paper: Amplified Analysis of DNA by the Autonomous Assembly of Polymers Consisting of DNAzyme Wires Fuan Wang, Johann Elbaz, Ron Orbach, Nimrod Magen and Itamar Willner*
More informationThr Gly Tyr. Gly Lys Asn
Your unique body characteristics (traits), such as hair color or blood type, are determined by the proteins your body produces. Proteins are the building blocks of life - in fact, about 45% of the human
More informationSupplementary information. USE1 is a bispecific conjugating enzyme for ubiquitin and FAT10. which FAT10ylates itself in cis
Supplementary information USE1 is a bispecific conjugating enzyme for ubiquitin and FAT10 which FAT10ylates itself in cis Annette Aichem 1, Christiane Pelzer 2, Sebastian Lukasiak 2, Birte Kalveram 2,
More information2.1 Calculate basic statistics
2.1 Calculate basic statistics The next part is an analysis performed on a FASTA formatted file containing complete genomic DNA (*.dna), not genes or proteins. Calculate the AT content (Per.AT), number
More informationInterpretation of sequence results
Interpretation of sequence results An overview on DNA sequencing: DNA sequencing involves the determination of the sequence of nucleotides in a sample of DNA. It use a modified PCR reaction where both
More informationSupplemental Data. Polymorphic Members of the lag Gene. Family Mediate Kin Discrimination. in Dictyostelium. Current Biology, Volume 19
Supplemental Data Polymorphic Members of the lag Gene Family Mediate Kin Discrimination in Dictyostelium Rocio Benabentos, Shigenori Hirose, Richard Sucgang, Tomaz Curk, Mariko Katoh, Elizabeth A. Ostrowski,
More informationThe HLA system. The Application of NGS to HLA Typing. Challenges in Data Interpretation
The Application of NGS to HLA Typing Challenges in Data Interpretation Marcelo A. Fernández Viña, Ph.D. Department of Pathology Medical School Stanford University The HLA system High degree of polymorphism
More informationCHAPTER II MATERIALS AND METHODS. Cell Culture and Plasmids. Cos-7 cells were maintained in Dulbecco s modified Eagle medium (DMEM,
CHAPTER II MATERIALS AND METHODS Cell Culture and Plasmids Cos-7 cells were maintained in Dulbecco s modified Eagle medium (DMEM, BioWhittaker Inc,Walkersville, MD) containing 10% fetal bovine serum, 50
More informationProtocol T45 Community Reference Laboratory for GM Food and Feed
EUROPEAN COMMISSION DIRECTORATE GENERAL JRC JOINT RESEARCH CENTRE INSTITUTE FOR HEALTH AND CONSUMER PROTECTION COMMUNITY REFERENCE LABORATORY FOR GM FOOD AND FEED Event-specific Method for the Quantification
More informationA suite of Gateway cloning vectors for high-throughput genetic analysis in Saccharomyces cerevisiae
Yeast Yeast 2007; 24: 913 919. Published online 21 June 2007 in Wiley InterScience (www.interscience.wiley.com).1502 Yeast Functional Analysis Report A suite of Gateway cloning vectors for high-throughput
More informationCloning and Expression of a Haloacid Dehalogenase Enzyme. By: Skyler Van Senior Research Advisor: Dr. Anne Roberts
Cloning and Expression of a Haloacid Dehalogenase Enzyme By: Skyler Van Senior Research Advisor: Dr. Anne Roberts utline The gene being cloned is JHP1130 from Helicobacter pylori (H. pylori) JHP1130 is
More informationNEBNext Multiplex Oligos for Illumina (Index Primers Set 2)
LIBRARY PREPARATION NEBNext Multiplex Oligos for Illumina (Index Primers Set 2) Instruction Manual NEB #E7500S/L 24/96 reactions Version 4.0 1/18 be INSPIRED drive DISCOVERY stay GENUINE This product is
More informationSupporting Online Material for
www.sciencemag.org/cgi/content/full/319/5867/1241/dc1 Supporting Online Material for Local Positive Feedback Regulation Determines Cell Shape in Root Hair Cells Seiji Takeda, Catherine Gapper, Hidetaka
More informationPLEASE SCROLL DOWN FOR ARTICLE
This article was downloaded by:[national Chung Hsing University] On: 8 November 2007 Access Details: [subscription number 770275792] Publisher: Informa Healthcare Informa Ltd Registered in England and
More informationRapid detection of a Dengue virus RNA sequence with single molecule sensitivity using tandem toehold-mediated displacement reactions
Electronic Supplementary Material (ESI) for ChemComm. This journal is The Royal Society of Chemistry 2018 Electronic Supplementary Information (ESI) for Rapid detection of a Dengue virus RNA sequence with
More informationAntigenic Variation of Ehrlichia chaffeensis Resulting from Differential Expression of the 28-Kilodalton Protein Gene Family
INFECTION AND IMMUNITY, Apr. 2002, p. 1824 1831 Vol. 70, No. 4 0019-9567/02/$04.00 0 DOI: 10.1128/IAI.70.4.1824 1831.2002 Copyright 2002, American Society for Microbiology. All Rights Reserved. Antigenic
More informationATTEMPT TO IDENTIFY NOVEL IFT MUTANT THROUGH PCR SEQUENCING AND ANALYSIS OF CHLAMYDOMONAS REINHARDTII FLAGELLAR MUTANTS
ATTEMPT TO IDENTIFY NOVEL IFT MUTANT THROUGH PCR SEQUENCING AND ANALYSIS OF CHLAMYDOMONAS REINHARDTII FLAGELLAR MUTANTS An Undergraduate Research Scholars Thesis by CATHERINE MARIE HERNANDEZ Submitted
More informationDiabetologia 9 Springer-Verlag 1992
Diabetologia (t 992) 35:743-747 Diabetologia 9 Springer-Verlag 1992 Human pancreatic Beta-cell glucokinase: cdna sequence and localization of the polymorphic gene to chromosome 7, band p 13 S. Nishi 1'3,
More informationpcmv-3tag Epitope Tagging Mammalian Expression Vectors
pcmv-3tag Epitope Tagging Mammalian Expression Vectors INSTRUCTION MANUAL Catalog #240195 (pcmv-3tag-1), #240196 (pcmv-3tag-2), #240197 (pcmv-3tag-3), #240198 (pcmv-3tag-4), #240200 (pcmv-3tag-6), #240202
More informationSupplementary Information. A chloroplast envelope bound PHD transcription factor mediates. chloroplast signals to the nucleus
Supplementary Information A chloroplast envelope bound PHD transcription factor mediates chloroplast signals to the nucleus Xuwu Sun, Peiqiang Feng, Xiumei Xu, Hailong Guo, Jinfang Ma, Wei Chi, Rongchen
More informationDRACULA2 is a dynamic nucleoporin with a role in regulating the shade. Marçal Gallemí, Anahit Galstyan, Sandi Paulišić, Christiane Then, Almudena
DRACULA2 is a dynamic nucleoporin with a role in regulating the shade avoidance syndrome in Arabidopsis. Marçal Gallemí, Anahit Galstyan, Sandi Paulišić, Christiane Then, Almudena Ferrández-Ayela, Laura
More informationUniversal Split Spinach Aptamer (USSA) for Nucleic Acid Analysis and DNA Computation
Electronic Supplementary Material (ESI) for ChemComm. This journal is The Royal Society of Chemistry 2017 Supporting materials Universal Split Spinach Aptamer (USSA) for Nucleic Acid Analysis and DNA Computation
More informationFour different segments of a DNA molecule are represented below.
Four different segments of a DNA molecule are represented below. There is an error in the DNA in which molecule? A. segment 1 only B. segment 3 only C. segment 2 and 3 D. segment 2 and 4 Explain the basic
More informationOlerup SSP KIR Genotyping
KIR Genotyping Product Insert Page 1 of 16 Olerup SSP KIR Genotyping Product number: Lot number: 98V Expiry date: 2016-December-01 Number of tests: 12 Number of wells per test: 23 + 1 Storage - pre-aliquoted
More informationPrimary structure of an extracellular matrix proteoglycan core protein deduced from cloned cdna
Proc. Natl. Acad. Sci. USA Vol. 83, pp. 7683-7687, October 1986 Biochemistry Primary structure of an extracellular matrix proteoglycan core protein deduced from cloned cdna TOM KRUSIUS AND ERKKI RUOSLAHTI
More informationElectronic Supplementary Information Sensitive detection of polynucleotide kinase using rolling circle amplification-induced chemiluminescence
Electronic Supplementary Material (ESI) for Chemical Communications. This journal is The Royal Society of Chemistry 2014 Electronic Supplementary Information Sensitive detection of polynucleotide kinase
More informationMolecular cloning of four novel murine ribonuclease genes: unusual expansion within the Ribonuclease A gene family
1997 Oxford University Press Nucleic Acids Research, 1997, Vol. 25, No. 21 4235 4239 Molecular cloning of four novel murine ribonuclease genes: unusual expansion within the Ribonuclease A gene family Dean
More informationSUPPLEMENTARY MATERIAL
SUPPLEMENTARY MATERIAL Addition of N ε -acetyl-lysine to the Genetic Code of Escherichia coli Heinz Neumann 1 Sew Y. Peak-Chew 1 & Jason W. Chin 1,2 1 Medical Research Council Laboratory of Molecular Biology,
More informationPorto, and ICBAS - Instituto de Ciências Biomédicas de Abel Salazar, Universidade do Porto, Porto, Portugal 2
Arch. Tierz., Dummerstorf 49 (2006) Special Issue, 103-108 1 CIIMAR - Centro Interdisciplinar de Investigação Marinha e Ambiental, R. dos Bragas, 177, 4050-123 Porto, and ICBAS - Instituto de Ciências
More informationMutations in the Human ATP-Binding Cassette Transporters ABCG5 and ABCG8 in Sitosterolemia
HUMAN MUTATION Mutation in Brief #518 (2002) Online MUTATION IN BRIEF Mutations in the Human ATP-Binding Cassette Transporters ABCG5 and ABCG8 in Sitosterolemia Susanne Heimerl 1, Thomas Langmann 1, Christoph
More informationEvent-specific Method for the Quantification of Soybean Line A Using Real-time PCR. Protocol
Event-specific Method for the Quantification of Soybean Line A2704-12 Using Real-time PCR Protocol 14 May 2007 Directorate General-Joint Research Centre Institute for Health and Consumer Protection Biotechnology
More informationProgrammable RNA microstructures for coordinated delivery of sirnas
Electronic Supplementary Material (ESI) for Nanoscale. This journal is The Royal Society of Chemistry 2016 Programmable RNA microstructures for coordinated delivery of sirnas Jaimie Marie Stewart, Mathias
More informationHuman Merkel Cell Polyomavirus Small T Antigen Is an Oncoprotein
Human Merkel Cell Polyomavirus Small T Antigen Is an Oncoprotein Targeting the 4EBP1 Translation Regulator Masahiro Shuda, Hyun Jin Kwun, Huichen Feng, Yuan Chang and Patrick S. Moore Supplemental data
More informationShoshan, David H. MacLennan, and Donald S. Wood, which appeared in the August 1981 issue ofproc. NatL Acad. Sci. USA
2124 Corrections Correction. n the article "Nucleotide sequence and the encoded amino acids of human serum albumin mrna" by Achilles Dugaiczyk, Simon W. Law, and Olivia E. Dennison, which appeared in the
More informationResearch Note. Evaluation of PCR-based approach for serotype determination of Streptococcus pneumoniae
Tropical Biomedicine 30(2): 338 344 (2013) Research Note Evaluation of PCR-based approach for serotype determination of Streptococcus pneumoniae Shakrin, N.N.S.M. 1, Balasubramaniam, S.D. 2, Yusof, H.A.
More informationUstilago maydis contains variable
The b mating-type locus of Ustilago maydis contains variable and constant regions James w. Kronstad t and Sally A. Leong 2 ~Biotechnology Laboratory, Departments of Microbiology and Plant Science, University
More informationCharacterization of the Azinomycin B Biosynthetic. Gene Cluster Revealing a Different Iterative Type I
Chemistry & Biology 15 Supplemental Data Characterization of the Azinomycin B Biosynthetic Gene Cluster Revealing a Different Iterative Type I Polyketide Synthase for Naphthoate Biosynthesis Qunfei Zhao,
More information1. DNA, RNA structure. 2. DNA replication. 3. Transcription, translation
1. DNA, RNA structure 2. DNA replication 3. Transcription, translation DNA and RNA are polymers of nucleotides DNA is a nucleic acid, made of long chains of nucleotides Nucleotide Phosphate group Nitrogenous
More informationMolecular Techniques Third-year Biology
PLANNING Genetics Lab practices Molecular Techniques. Genetics Lab practices protocol. 2015-16 PCR-DIRECTED MUTAGENESIS, MOLECULAR CLONING AND RESTRICTION ANALYSIS Sessions 1 & 2 (2x3 hours): PCR-directed
More informationRibozymes and sirna Protocols
METHODS IN MOLECULAR BIOLOGY TM TM Volume 252 Ribozymes and sirna Protocols SECOND EDITION Edited by Mouldy Sioud pred GFP pred GFP sirna strsv-derived Hairpin Ribozymes 339 25 Design, Targeting, and Initial
More informationNucleic Acids Research
Volume 14 Number 16 1986 Nucleic Acids Research Nucleotide sequence of the Eschenchia coli replication gene dnazx Kuo-Chang Yin, Aleksandra Blinkowa and James R.Walker Department of Microbiology, University
More informationA Conserved -Helix Essential for a Type VI Secretion-Like System of Francisella tularensis
JOURNAL OF BACTERIOLOGY, Apr. 2009, p. 2431 2446 Vol. 191, No. 8 0021-9193/09/$08.00 0 doi:10.1128/jb.01759-08 Copyright 2009, American Society for Microbiology. All Rights Reserved. A Conserved -Helix
More informationChemical synthesis of the thymidylate synthase gene
roc. Natl. Acad. Sci. USA Vol. 87, pp. 633-637, January 1990 Biochemistry Chemical synthesis of the thymidylate synthase gene (gene synthesis/cassette mutagenesis/protein engineering) SHANE CLIMIE AND
More informationTRANSPOSON INSERTION SITE VERIFICATION
TRANSPOSON INSERTION SITE VERIFICATION Transposon and T-DNA insertion in Arabidopsis genes can be identified using the Arabidopsis thaliana Insertion Database (ATIdb) (http://atidb.org/cgi-perl/gbrowse/atibrowse).
More informationGateway Vectors for BiFC
Gateway Vectors for BiFC 1. The enhanced YFP (EYFP) are used (Split EYFP). 2. The Fusion fusion gene is expressed by CaMV35S promoter. 3. The N- or C-terminal fragments of EYFP are fused subsequent to
More informationTA Cloning Kit. Catalog nos. K and K Catalog nos. K and K Catalog nos. K and K
TA Cloning Kit Version T August 20, 2002 25-0024 TA Cloning Kit Catalog nos. K2000-01 and K2000-40 Catalog nos. K2030-01 and K2030-40 Catalog nos. K2040-01 and K2040-40 A Limited Label License covers this
More informationProtein Synthesis: Transcription and Translation
Review Protein Synthesis: Transcription and Translation Central Dogma of Molecular Biology Protein synthesis requires two steps: transcription and translation. DNA contains codes Three bases in DNA code
More informationGENETICS and the DNA code NOTES
GENETICS and the DNA code NOTES BACKGROUND DNA is the hereditary material of most organisms. It is an organic compound made of two strands, twisted around one another to form a double helix. Each strand
More informationBiomolecules: lecture 6
Biomolecules: lecture 6 - to learn the basics on how DNA serves to make RNA = transcription - to learn how the genetic code instructs protein synthesis - to learn the basics on how proteins are synthesized
More informationProtein Synthesis. Application Based Questions
Protein Synthesis Application Based Questions MRNA Triplet Codons Note: Logic behind the single letter abbreviations can be found at: http://www.biology.arizona.edu/biochemistry/problem_sets/aa/dayhoff.html
More informationSTUDIES ON THE FUSION OF LIGNINOLYTIC ENZYME cdnas AND THEIR EXPRESSION
STUDIES ON THE FUSION OF LIGNINOLYTIC ENZYME cdnas AND THEIR EXPRESSION Jun Xie, a* Lei Feng, a Ning Xu, a Guohui Zhu, a Jun Yang, a Xiaoli Xu, a and Shiyu Fu b* Manganese peroxidase (MnP) and lignin peroxidase
More informationComplete mitochondrial DNA sequence of the Japanese eel Anguilla japonica
FISHERIES SCIENCE 2001; 67: 118 125 Original Article Complete mitochondrial DNA sequence of the Japanese eel Anguilla japonica JUN G INOUE, 1, * MASAKI MIYA, 2 JUN AOYAMA, 1 SATOSHI ISHIKAWA, 1 KATSUMI
More information