Blue Heron, Your Gene Synthesis Partner You Design it We Build it Simple to Complex Sequences Codon Optimization Any species Variants Single or pooled clone libraries Antibody Affinity Optimization Whole Genome Synthesis Custom Vector Construction
Blue Heron Gene Synthesis A Pioneer and provider of the highest quality and most comprehensive custom gene synthesis services to researchers worldwide since 1999. By working with Blue Heron you save money and valuable time by leaving the synthesis work to the experts. You receive all your genes from the simplest to the most complex in your vector of choice, on time, and with no surprises. Unlike approaches that rely on adaptations of standard PCR, our GeneMaker platforms include exclusive design software that considers multiple characteristic of each section of the target DNA sequence, and optimally builds each section using multiple, proprietary process technologies. This way, Blue Heron successfully completes projects where our competitors' processes often fail. Researchers from the top 20 pharmaceutical and biotechnology companies, as well as emerging synthetic biology and biofuels companies like The Venter Institute; choose Blue Heron for our unmatched technological capability, quality of customer service, and long record of success and industry leadership. PARTNERS IN DISCOVERY Gene synthesis provides customers with the most cost effective and reliable source to obtain genes that are not already available from cloned libraries. It also allows researchers to build novel variant libraries or entire genomes that can lead to new discoveries for the protein, antibody discovery, synthetic biology and biofuels industries. RESEARCH BENEFITS n Get any gene or synthetic DNA sequence n Free codon optimization- any species n Expression vector cloning n Secure and confidential data transfer n No claims to your IP QUALITY n Automated ordering process to minimize errors n Unmatched technology platforms n 2x sequence verified data confirmation n Technical customer service TECHNOLOGY LEADER n 52kb length gene delivered n First megabase a month capacity n First synthetic genome partnership n Most complex sequences synthesized RESEARCH ARTICLE Creation of a Bacterial Cell Controlled by a Chemically Synthesized Genome Daniel G. Gibson 1, John I. Glass 1, Carole Lartigue 1, Vladimir N. Noskov 1, Ray-Yuan Chuang 1, Mikkel A. Algire 1, Gwynedd A. Benders 2, Michael G. Montague 1, Li Ma 1, Monzia M. Moodie 1, Chuck Merryman 1, Sanjay Vashee 1, Radha Krishnakumar 1, Nacyra Assad-Garcia 1, Cynthia Andrews-Pfannkoch 1, Evgeniya A. Denisova 1, Lei Young 1, Zhi-Qing Qi 1, Thomas H. Segall-Shapiro 1, Christopher H. Calvey 1, Prashanth P. Parmar 1, Clyde A. Hutchison III 2, Hamilton O. Smith 2 and J. Craig Venter 1,2, * UNMATCHED SERVICE n Confidentiality guaranteed n On time delivery n Customer service ready to help n Automated quoting tools n Quotes to orders with a simple click Science 329, 52-56. July, 2010. www.blueheronbio.com
Cloning Services Once a gene or synthetic DNA fragment has been synthesized, Blue Heron offers standard vector as well as custom expression vector cloning. Using our custom expression vector services provides you with a finished, expression ready product for your downstream projects. RESEARCH BENEFITS Plasmid Prep Services MIDI AND MAXI-PREP n Ion exchange column purified n Receive transfection-grade DNA n Sequence verified n Receive the quantity you need 100ug 500ug 1000ug n Cloned and ready for the next step n Sequence verified in the final vector APPLICATIONS n Protein expression n Antibody development n shrna or rare RNA template n Transgenics studies CUSTOM VECTOR DESIGN n Reshuffled vector sites n Multi-resistance vector n Specialized cassette drop-in vectors n Custom designed vectors ORIGENE EXPRESSION VECTORS n Mammalian expression n E.coli expression n Epitope and fluorescent tags 425-368-5000
Generating Diversity- Synthetic Variants Antibody Engineering to Increase Affinity Creating diversity through random mutagenesis, recombination or degenerate oligo methods can be cumbersome as well as highly inaccurate. Using Blue Heron s gene synthesis technology to create synthetic variants enables a researcher the design flexibility and guarantee of sequence needed. Light Chain (L) Antigen Antigen-binding site Heavy Chain (H) C H C L VH C = Constant Region V = Constant Region Complementary Determining Region (CDR) VL BENEFITS n Study multiple variants in parallel n Receive ready for protein expression n Available as individual clones or as a pool n High quality pools decrease downstream screening costs QUALITY n Sequence verified n Cost/time line defined APPLICATIONS n Mutation analysis n Antibody discovery n Gene/protein function studies types n Simple to complex base variants n Amino acid substitutions n Domain swaps n Heavily mutated promoter regions n Fusion or tagged proteins n Knockout constructs n Deletions/insertions n Alternative splice forms PROCESS n You submit the full CDR plus variable region DNA sequence and the expression vector. n We create a custom vector containing the constant regions specific for your project. n We build the variants, insert the variable region cassettes into the custom vector. n Deliverable: Full-length, sequence-verified, individual antibody clones in your expression vector. ADVANTAGES n Flexibility to make any amino acid change within the variable region cassette. n Delivery reliability n Faster than using traditional mutagenesis methods Cassette Variant Design New Vector Containing Constant Region(s) Constant Region 1 Constant Region 2 Variant 1 Variant 2 Variant 3 Variant 4 Your variants in the New Vector containing constant region(s) www.blueheronbio.com
Codon Optimization- Free Tool Blue Heron provides customers with a free Codon Optimization tool to assist in sequence design. In many cases researchers have found improved protein expression results after optimization (see below). Each optimization request generates an optimized sequence posted electronically to the customer's dedicated, secure website. RESEARCH BENEFITS n Improve protein expression n Heterologous expression improvement n Remove specific DNA sequences or motifs n Diminish complexity at the DNA level LGR4 AND LGR6 PROTEIN EXPRESSION IMPROVED USING BLUE HERON'S CODON OPTIMIZATION ALGORITHM 1 2 3 4 Abbreviation (3 Letter) Full Name Codon options Alanine Ala GCT GCC GCA GCG Arginine Arg CGT CGC CGA CGG AGA AGG Asparagine Asn AAT AAC Aspartate Asp GAT GAC Cysteine Cys TGT TGC Glutamate Glu GAA GAG CAA CAG Glycine Gly GGT GGC GGA GGG Histidine His CAT CAC Isoleucine Ile ATT ATC ATA Leucine Leu TTA TTG CTT CTC CTA CTG 116 116 Lysine Lys AAA AAG 66 45 35 25 LGR4 66 45 35 25 LGR6 Methionine Met ATG Phenylalanine Phe TTT TTC 18 14 18 14 Proline Pro CCT CCC CCA CCG Lane 1: construct with a wild type LGR4 DNA sequence Lane 2: construct with optimized LGR4 DNA sequence Lane 3: construct with a wild type LGR6 DNA sequence Lane 4: construct with optimized LGR6 DNA sequence Protocol: LGR4 and LGR6 proteins were codon optimized using Blue Heron s proprietary algorithm. Wild type and optimized sequences were cloned separately into OriGene s spcmv-entry vector. Each DNA was transiently transfected to HEK293T cells and incubated for 72 hrs. Proteins were purified using anti-ddk affinitive column and run on SDS PAGE, then stained with Coomassie Blue. Serine Ser TCT TCC TCA TCG AGT AGC Threonine Thr ACT ACC ACA ACG Tryptophan Trp TGG Tyrosine Tyr TAT TAC Valine Val GTT GTC GTA GTG 425-368-5000
Automated Design Tools n Project design tool NEW! n Automated quote generation n Automated quotes to orders NEW! n Free codon optimization & design n Secure, confidential ordering web site n Set up expression vectors for cloning n Choose to have a midi or maxi prep prepared n Sequence and trace data automatically posted DELIVERABLES: n 3-5 μg purified plasmid DNA Bacterial strain containing the plasmid n Sequencing data and trace data posted electronically to your secure, dedicated online account PROJECT DESIGN TOOL Ordering is fast and simple: Step 1 Step 2 Step 3 Enter the DNA Sequence you need Choose your vector (Standard or Expression) Check your shipping/billing details DONE! n www.blueheronbio.com PROJECT VERIFICATION PAGE
Molecular and Protein Biology Solutions TrueORF/Gold Human/mouse Expression vectors 5,000 humn Custom purification TrueMAB TruePlex Clones/genes Proteins Antibodies Assays Any species Any sequence Any variant Optimized expression Antibody engineering As of August, 2010, Blue Heron became a wholly owned subsidiary of OriGene Technologies, Inc. Combining OriGene s complete collection of human cdna clones with Blue Heron s gene synthesis capacity, we can provide a whole product solution for the molecular biology research community. We are 100% committed to serving our customers with high quality, strict confidentiality standards, and improved business efficiency OriGene Product Line GENE EXPRESSION n TissueScan cancer qpcr arrays n Genome wide qpcr primer pairs n qpcr template standards n microrna qpcr detection system PROTEINS AND ANTIBODIES n 5,000 purified human proteins n Heavy-labeled proteins as MS standard n 12,000 transfected over-expression lysates n TrueMAB monoclonal antibodies CDNA CLONES: OVER-EXPRESSION n TrueORF : tagged cdna clones n TrueORF gold clones: expression validated n Over 50 organelle markers n TrueClone : native protein expression n microrna expression plasmids RNAi: DOWN-REGULATION n Pre-designed HuSH-29 shrna n Every human, mouse and rat genes n Guaranteed knock down n microrna expression plasmids 425-368-5000
OriGene, Your Partner in Gene Research and Beyond KEY TECHNOLOGIES AND PRODUCTS n Full-length cdna clones, ORF clones in expression-ready vectors n Gene synthesis: any gene, any variant, any vector n RNAi research reagents: shrna, sirna, and mirna function and detection n SYBR Green qpcr assays for mrna and mirna detection, primer panels n Recombinant human proteins n TrueMAB monoclonal antibodies n Luminex multiplex immunoassays n Cancer tissue biorepository, TMA, RPPA, and TissueScan qpcr arrays Blue Heron-Gene Synthesis Headquarters: Bothell, WA USA Contact Us: www.blueheronbio.com 1-425-368-5000 Sales: sales@blueheronbio.com Customer Service: customerservice@blueheronbio.com OriGene Technologies, Inc.: Rockville, MD USA Contact Us: www.origene.com 1-301-340-3188 1-888-267-4436 (U.S. only) Customersupport@origene.com Techsupport@origene.com