Adam Clore presents on IDT's gBlocks Gene Fragments, which are synthetic linear DNA fragments up to 2000 base pairs in length that are high fidelity and sequence verified. Some key applications of gBlocks Gene Fragments include gene and vector construction, metabolic engineering, viral and vaccine development, protein evolution, next generation sequencing controls, antibody expression, PCR controls, CRISPR/Cas9 genome editing, and homology directed repair. IDT offers design support and online tools to assist customers with ordering and designing gBlocks Gene Fragments.

1. 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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2. gBlocks® Gene Fragments
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3. 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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4. High sequence fidelity of gBlocks® Gene Fragments
Data generated on Illumina sequencing platforms from 262 customer
gBlocks Gene Fragments

5. gBlocks® Gene Fragments
applications
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6. gBlocks® Gene Fragments are used for a variety of applications
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>100 citations available at www.idtdna.com/gBlocks

7. 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 amplify
Key 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

8. 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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9. 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

10. 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

11. 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

12. 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

13. 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 Rettberg
President, iGEM Foundation

14. New tools for researchers
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15. 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

16. 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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17. • Shows item name, progress, and
estimated and actual ship dates
• Quickly download QC documents
Online tools—order status page
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18. 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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19. Rewrite of rules with dynamic scoring
– Scores reflect the severity of
the complexity
Improved rules engine for sequence acceptance
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20. • Single stranded gBlocks
Gene Fragments for
homology-mediated repair
• Up to 2.5 kb
In beta testing now
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21. 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
genes@idtdna.com
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22. 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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23. • Support for custom gene synthesis and
gBlocks® Gene Fragments, including libraries:
genes@idtdna.com
• 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

24. Questions?
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