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Adam Clore, PhD
Gene synthesis technology and applications update—unleash your lab’s potential with IDT Genes and gBlocks® Gene Fragments
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gBlocks® Gene Fragments
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The first commercially available synthetic, linear, double-stranded DNA fragments:• High fidelity and sequence-verified• Complete custom flexibility
for a variety of applications• Lengths up to 2000 bp• Yields up to 1 µg• Libraries available • Fast delivery—median shipping time
of just 3 business days
What are gBlocks® Gene Fragments?
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High sequence fidelity of gBlocks® Gene Fragments
Data generated on Illumina sequencing platforms from 262 customer gBlocks Gene Fragments
gBlocks® Gene Fragments applications
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gBlocks® Gene Fragments are used for a variety of applications
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>100 citations available at www.idtdna.com/gBlocks
Building genes everything– Genes/routine cloning– Metabolic engineering – Viruses/vaccines– Protein evolution
Excellent alternative to PCR and restriction digest methods
– Less time and effort to optimize – Get the sequences that you want,
not just what you can amplifyKey benefits
– Sequence acceptance criteria– Constancy in TAT and minimum delays
How can we help?– Codon optimization– Split large sequences into
gBlocks Gene Fragments– Order tracking tool
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gBlocks® Gene Fragments applications
NGS controls – Spike-ins to verify pull down
efficiency– Spike-ins for sample tracking
Linear DNA – No plasmid backbones– No genomic DNA contamination
Key benefits– Flexible sequence design– Lack of cross contamination
How can we help?– Sequence design options– Contamination data from NGS– Plates and stamping
gBlocks® Gene Fragments applications
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Antibody expression – CAR T-cell therapy– Antibody-based
vaccines/therapies
“How much time and money do you spend screening antibody libraries?”
– Spend it doing research, not cloning!
Key benefits– Large numbers of sequences to
screen (custom libraries) – High throughput (plates, volume)– Low price per construct
How can we help?– Library design assistance – Plate orders – Sequence optimization
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gBlocks® Gene Fragments applications
PCR control templates – Diagnostics– Detection assays
Key benefits– No cross contamination– Sequence accuracy – Custom formulation
How can we help?– Design buffer sequences
needed to increase length and manufacturing reliability
Do or will you need GMP controls?– IDT offers GMP Genes
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gBlocks® Gene Fragments applications
CRISPR guide RNAs and gene constructs
gBlocks Gene Fragments have been used in numerous published CRISPR studies:
– Generating Cas9 and sgRNA vectors– Transfecting sgRNA expression
cassettes under U6 promoter control as described by the Church Lab (Harvard Medical School, Boston MA)
Citations at www.idtdna.com/gBlocks
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For genome editing, we recommend the Alt-R™ CRISPR-Cas9 System:
www.idtdna.com/CRISPR.
gBlocks® Gene Fragments applications
Donor DNA for homology directed repair
– Knock ins
Key features– Sequence acceptance criteria– TAT
How can we help?– Split large sequences into
gBlocks Gene Fragments– Homology arm design with
Alt-R™ CRISPR crRNA design
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gBlocks® Gene Fragments applications
2016 IDT iGEM sponsorship
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20 kb of DNA as gBlocks® Gene Fragments free of charge to each iGEM 2016 team!• 232 iGEM teams have signed up for the
free DNA offer this year
• Registration at www.idtdna.com/iGEM
“Thanks to IDT, the teams can focus on imagining, engineering, and testing the novel devices they create to solve important world problems.”
—Randy RettbergPresident, iGEM Foundation
New tools for researchers
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Complexities within sequences can be highlighted on the ordering site
– Sequences can be edited without leaving the ordering tool
Simple codon optimization exists within the gBlocks Gene Fragments ordering tool
Online tools—editing tools
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YouTube tutorials are available for both complexities and codon optimization
The stand-alone codon optimization tool contains additional functionality outside of the ordering pages
A YouTube tutorial is available for the stand-‐alone codon optimization tool
Online tools—codon optimization
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• Shows item name, progress, and estimated and actual ship dates
• Quickly download QC documents
Online tools—order status page
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Currently for genes >500 bp– NGS data is more accurate than
Sanger sequencing– Sequence data feeds into the
rules engine updates
NGS analysis of Genes
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Rewrite of rules with dynamic scoring– Scores reflect the severity of
the complexity
Improved rules engine for sequence acceptance
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• Single stranded gBlocks Gene Fragments for homology-mediated repair
• Up to 2.5 kb
In beta testing now
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Free custom design support for gBlocks® Gene Fragments
• Do you need help with a unique design?– Examples of custom requests we have assisted with:
• 1900 bp sequence: no C bases on one strand and no G bases on the other
• Generate codon usage tables given unique sequencing data• Minimize CpG dinucleotides, but simultaneously maximize G/C in the 3rd
codon position
• Do you have other custom requests?– For more information about IDT gene synthesis products, email
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We screen the sequence of every Gene and gBlocks® Gene Fragment ordered for potentially dangerous sequences:• IDT is a founding member of the
International Gene Synthesis Consortium (IGSC)
• We ensure safety and regulatory conformance
• For more information, visit www.genesynthesisconsortium.org
Biosecurity is important at IDT
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• Support for custom gene synthesis and gBlocks® Gene Fragments, including libraries: [email protected]
• gBlocks Gene Fragments information, citations, and ordering: www.idtdna.com/gBlocks
• Custom gene synthesis information and ordering: www.idtdna.com/genes
Support and additional resources
Questions?
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