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gBlocks Gene Fragments

Pure confidence

gBlocks HiFi Gene Fragments (1000–3000 bp in length) extend our offerings of double-stranded fragments, and are optimized for the assembly of large constructs. Pick fewer colonies and drive your projects to completion faster with these high purity fragments.

NEW gBlocks HiFi Gene Fragments (1000–3000 bp in length) extend our offerings of double-stranded fragments, and are optimized for the assembly of large constructs. Pick fewer colonies and drive your projects to completion faster with these high purity fragments.

  • High fidelity and purity for gene assembly
  • Compatible with all downstream cloning methods
  • No universal adapters to remove

Ordering

gBlocks gene fragments in tubes

A, T, C, and G residues only. Delivered dry and normalized to 250, 500, or 1000 ng, depending on length.

gBlocks gene fragments in plates

Available for fragments of 125–3000 bp. Orders require a minimum of 24 fragments per plate. Resuspended in 25 μL of nuclease-free water (concentration: 10 ng/μL). Shipped on dry ice and delivered within 10 business days of order confirmation (excluding Fridays). See below for pricing, per fragment, based on fragment length.


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gBlocks HiFi Gene Fragments in tubes

There are no order minimums for tubes. Shipped on dry ice and shipped within 5–8 business days of order confirmation (excluding Fridays).

gBlocks HiFi Gene Fragments in plates

Orders require a minimum of 24 fragments per plate. Resuspended in 25 μL of nuclease-free water (concentration: 10 ng/μL). Shipped on dry ice and delivered within 10 business days of order confirmation (excluding Fridays).

gBlocks Gene Fragment libraries in tubes

Delivered dry, normalized to 200 ng. Libraries are not available in plate format.

1 BD: business days. Shipping time is dependent on length and complexity of the gBlocks fragments ordered. In a few cases, shipping time may exceed the estimated time.

Sequence Information is secure and confidential at IDT. Please see our Confidentiality Statement for more information.

gBlocks Gene Fragments are sequence-verified, double-stranded DNA molecules 125–3000 bp in length. They are manufactured with the same industry-leading, high-fidelity synthesis chemistries that were developed for our Ultramer DNA Oligos. Their outstanding quality and rapid delivery time make them ideal for quick gene construction or modification, and any other application requiring double-stranded DNA. These include, but are not limited to:

  • CRISPR-based genome editing
  • Recombinant antibody engineering
  • qPCR and PCR controls
  • Enzyme engineering
  • Vaccine research
  • High resolution melt (HRM) assays

gBlocks Gene Fragments

gBlocks Gene Fragments are double-stranded DNA fragments 125–3000 bp in length. They are manufactured with the same industry-leading, high-fidelity synthesis chemistries that were developed for our Ultramer DNA Oligos. gBlocks are available with standard A, C, G, T bases as well as up to 18 consecutive N or K residues.

Each gBlocks Gene Fragment goes through a quality control and sequence verification process. This includes size verification by capillary electrophoresis and sequence identification by mass spectrometry. This rigorous testing ensures that most recombinant colonies obtained from cloning each gBlocks Gene Fragment will contain the desired insert. More complex sequences may need the end user to sequence additional clones.

NEW gBlocks HiFi Gene Fragments

gBlocks HiFi Gene Fragments are double-stranded DNA fragments with sizes between 1000-3000 bp and verified with an error rate of less than 1:12,000 via NGS. These high-quality, high-fidelity constructs facilitate the assembly of large and complex sequences, matching both the length and accuracy needed to minimize the introduction of unwanted substitution or deletion errors.

With either gBlocks or gBlocks HiFi Gene Fragments, you can easily assemble and clone your DNA fragment into the vector of your choice using a variety of cloning methods, including the Gibson Assembly® method and blunt- or cohesive-end cloning protocols. For added flexibility, you can order gBlocks Gene Fragments with or without a 5′-phosphate group.

gBlocks Gene Fragment Libraries

gBlocks Gene Fragment Libraries are pooled gBlocks fragments of 251 to 500 bp in total length that are ideal for generating recombinant antibodies or for protein engineering. With gBlocks Gene Fragments Libraries, you can generate hundreds of thousands of constructs at a fraction of the cost that creating multiple variant libraries would require. The variable regions can be up to 18 consecutive N or K bases long and must be at least 125 bp from either end of the gene fragment (Figure 1).

Figure 1. Ordering format for gBlocks Gene Fragment Libraries. Placing a K mixed base in the third position of codons eliminates the TAA and TGA stop codons from being included in the gene fragments libraries, leaving only TAG as the possible stop codon.

Need something else? Tell us about it! We realize that we have a limited offering for gBlocks Gene Fragments Libraries. We will be offering more complex libraries in the future. Help us prioritize our product development by sharing what you need from us with as much detail as possible (i.e., show us sequences, including features such as mixed bases, annotations, and drawings) and send to libraries@idtdna.com.

Why do we limit the variable regions to 18 mixed bases?

When you order a gene fragment library that contains 18 "N" mixed bases, you order a pool of 418 gene fragments—about 68.7 billion different gBlocks Gene Fragments. Increasing the number of variable bases beyond 18 would result in more sequence combinations than can be accommodated in the 200 ng of product provided, which would compromise the overall representation of sequences within the pool.

Robust and flexible assembly

gBlocks Gene Fragments can be easily assembled and cloned into the vector of your choice using a variety of cloning methods, including the Gibson Assembly® method [1] and blunt- or cohesive-end cloning protocols. The fidelity of gBlocks fragments substantially reduces the number of clones that need to be sequenced to identify the intended construct when compared to other methods of gene assembly using DNA oligonucleotides. Note that assembly methods such as blunt-end cloning and restriction cloning consistently demonstrate lower fidelity. For added flexibility, you can order gBlocks Gene Fragments with or without a 5′-phosphate group

Adding diversity at reasonable cost

For screening purposes, gBlocks Gene Fragments Libraries allow for the generation of hundreds of thousands of constructs at a fraction of the cost of generating variant libraries.

References

  1. Gibson D, Young L, et al. (2009) Enzymatic assembly of DNA molecules up to several hundred kilobases. Nat Methods, 6(5):343–345.

Quality assurance

Every gBlocks Gene Fragment you receive goes through the following verification process:

  1. Sequences (125–3000 base pairs) are entered on the gBlocks Gene Fragments order entry page. A, C, G, T bases, as well as up to 18 consecutive N or K variable bases can be entered. Other mixed degenerate bases are currently not available, and neither are “modified" bases (inosine, uridine).
  2. Upon order entry, the entered sequences are reviewed by both our automated screening tools and our scientists to identify characteristics that may interfere with synthesis.
  3. Biosafety and regulatory conformance check:
    • A biohazard disclosure statement is required for each gene order.
    • IDT reserves the right to refuse any order that does not pass this analysis. If one of your sequences does not pass the screen criteria, you will be contacted by a gene services specialist to determine the best way to proceed.

Quality control and sequence verification

Each gBlocks Gene Fragment undergoes the following quality control procedures:

  1. Size verification by capillary electrophoresis
  2. Sequence identification by mass spectrometry

This rigorous testing ensures that, in the majority of cases, >80% of recombinant colonies obtained from cloning each gBlocks Gene Fragment will contain the desired insert. More complex sequences may suffer from reduced fidelity, which can be addressed by the end user sequencing additional clones.

For gBlocks Gene Fragment Libraries, each constant region is verified as above. The final library product is size verified by capillary electrophoresis.

High fidelity and purity for gene assembly

Both gBlocks and gBlocks HiFi Gene Fragments demonstrate consistent high sequence fidelity and purity across various lengths.

IDT gene fragments are compatible with all cloning methods that require double-stranded DNA as a starting material, allowing easy assembly of your desired construct sequence into your favorite cloning system. Compatible cloning systems include but are not limited to traditional cloning, Gibson Assembly, Golden Gate, Gateway, TOPO/TA cloning, and Blunt End cloning.

Figure 2: The effect of error rate on predicted cloning success. With an industry leading error rate of 1:12,000, gBlocks HiFi Gene Fragments demonstrate a high probability of first-time cloning success. Compared with other leading suppliers, gBlocks HiFi Gene Fragments are ~45% more likely to give a correct clone the first time when cloning fragments between 1000 and 3000 bps. Standard gBlocks Gene Fragments are still an excellent option for smaller fragment length ranges under 1000 bp.
Figure 3: Demonstrated cloning efficiency. Based on Illumina NGS sequencing of approximately 500 clones, gBlocks HiFi Gene Fragments exhibited a high degree of cloning success. Compared to a leading provider, gBlocks HiFi fragments showed a significant improvement in cloning efficiency leading to a reduction in the time and cost to find a correct clone. gBlocks Gene Fragments with lengths of 223-296 bp were cloned via restriction cloning and sequence verified to determine cloning efficiency.

Table 1 demonstrates the approximate number of colonies needed to be picked to have a >90% chance of obtaining a desired clone.

Table 1. Minimal screening effort is needed with gBlocks and gBlocks HiFi Gene Fragments to find a correct colony

Length (bp)gBlocks Gene FragmentsgBlocks HiFi Gene FragmentsOther Supplier
≤1000 3N/A5
1001-2000326-10
2001-300042N/A

 

* Single gBlocks Gene Fragments ranging between 223 and 296 bp were cloned into pUC57, pBluescript II, pET27, psiCHECK-2, Zero Blunt TOPO, pIDTSMART, and pGEM T Easy plasmids. Resulting clones were sequence verified by double-stranded sequencing.

User guides and protocols

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Frequently asked questions

Sequence Information is secure and confidential at IDT. Please see our Confidentiality Statement for more information. All online ordering steps, including sequence entry and your choice of parameters, are also secure and protected.

We screen the sequence of every gene fragment order we receive to (1) identify any regulated and other potentially dangerous pathogen sequences, and (2) verify that IDT’s gene customers are legitimate scientists engaged in beneficial research.

IDT is among the five founding members of the International Gene Synthesis Consortium (IGSC) and helped to create the IGSC’s Harmonized Screening Protocol. The Harmonized Screening Protocol describes the gene sequence and customer screening practices that IGSC member companies employ to prevent the misuse of synthetic genes. IDT takes the steps set out in the Harmonized Screening Protocol to screen the sequences of ordered genes and the prospective customers who submit those orders.

For more information about the IGSC and the Harmonized Screening Protocol, please visit the website at www.genesynthesisconsortium.org.

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In October 2010, the United States government issued final Screening Framework Guidance for Providers of Synthetic Double-Stranded DNA, describing how commercial providers of synthetic genes should perform gene sequence and customer screening. IDT and the other IGSC member companies supported the adoption of the Screening Framework Guidance, and IDT follows that Guidance in its application of the Harmonized Screening Protocol. For more information, please see 75 FR 62820 (Oct. 13, 2010), or https://federalregister.gov/a/2010-25728.

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