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Easy gene assembly—gBlocks Gene Fragments

Could your research benefit from faster gene assembly? gBlocks® Gene Fragments are 125–3000 bp, custom double-stranded DNA fragments of your specified sequence. Use them for all your cloning applications.

Apr 11, 2012

Revised/updated Nov 17, 2016

  • Highly versatile—These double-stranded DNA fragments of up to 3 kb can be used to easily and reliably assemble almost any sequence, and are compatible with most published cloning methods, including the Gibson Assembly® Method, and blunt-end or cohesive-end cloning protocols.
  • Easy isothermal gene assembly—Using the Gibson Assembly Method, multiple gBlocks Gene Fragments can be assembled into a larger gene construct, in a single reaction that takes about 1 hour. A simple, 20–80 nt sequence overlap is required when designing the gene fragments for assembly.
  • Affordable—gBlocks Gene Fragments are up to half the price of other synthetic gene constructs, making synthetic biology accessible to any lab.
  • Short delivery time—gBlocks Gene Fragments are typically shipped within 5–8 business days.
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Figure 1. gBlocks Gene Fragments make it easy to produce the correct synthetic genes. gBlocks Gene Fragments (n=43), ranging from 126 to 459 bp and with GC ratios between 40 and 70%, were synthesized and cloned into pIDTSMART-Amp by blunt-end cloning. Ligated plasmids were transformed into XL1Blue cells, and 3–25 clones for each gBlocks Gene Fragment were selected and sequenced using traditional Sanger sequencing. The average fidelity for all sequences analyzed was 90%, and for 40 out of 43 gBlocks Gene Fragments the correct sequence was observed in >80% clones. The high percentage of correct sequences obtained makes it easy to find your desired clone.

  

High-fidelity, double-stranded DNA sequences

gBlocks Gene Fragments are sequence-verified, double-stranded DNA fragments, 125–3000 bp in length, that enable easy gene construction or modification. They are synthesized using the highest fidelity synthesis methods, developed by IDT for our industry-leading Ultramer® Oligonucleotides, making them ideal for a broad range of cloning techniques.

Since they were introduced in 2012, researchers have been using gBlocks Gene Fragments for many unique applications. The Church laboratory at Harvard University (Boston, MA, USA) published a protocol for designing gRNAs from gBlocks Gene Fragments, and transferring these gRNAs and Cas9 constructs into human-induced pluripotent stem cells and HEK293 cells [1]. gBlocks Gene Fragments and the Gibson Assembly® method [2] can also be used as a reliable method for construct assembly when commercial clones to a poorly characterized gene are unavailable, such as in the case of the Micalcl protein, as described by Marbiah MM, Harvey A, et al [3].

Screen fewer clones

Experimental data shows that 90% of cloned gBlocks Gene Fragments will have the correct sequence (Figure 1).

gBlocks Fidelity

Figure 2. gBlocks Gene Fragments make it easy to produce the correct synthetic genes. gBlocks Gene Fragments (n=43), ranging from 126 to 459 bp and with GC ratios between 40 and 70%, were synthesized and cloned into pIDTSMART-Amp by blunt-end cloning. Ligated plasmids were transformed into XL1Blue cells, and 3–25 clones for each gBlocks Gene Fragment were selected and sequenced using traditional Sanger sequencing. The average fidelity for all sequences analyzed was 90%, and for 40 out of 43 gBlocks Gene Fragments the correct sequence was observed in >80% clones. The high percentage of correct sequences obtained makes it easy to find your desired clone.

In addition, using the Gibson Assembly method, two or more gBlocks fragments can easily be assembled in a single reaction to generate larger constructs [2] (Table 1). gBlocks Gene Fragments are synthesized without a 5′ phosphate; optional 5′ phosphorylation is available for blunt cloning.

Table 1. Isothermal Assembly of Multiple gBlocks Gene Fragments. EGFP and Kanamycin genes were assembled from 2 and 3 gBlocks Gene Fragments respectively and cloned into a linearized pUC57 vector using the Gibson Assembly method [2]. Cloned sequences were verified by double-stranded Sanger sequencing. The results show that even when assembling 3 fragments, gBlocks Gene Fragments make gene construction a highly reliable process.
Wild-type clones Gene length (nt) gBlocks Gene Fragments (#) Correct clones
EFGP 720 2 3/8
Kanamycin 816 3 2/8

Get gBlocks Gene Fragments quickly

The easy online ordering system allows sequences for gBlocks Gene Fragments to be entered individually or as multiple entries using a provided Microsoft Excel template. Orders typically ship within 5–8 business days. gBlocks Gene Fragments are delivered with a short protocol that summarizes cloning methods for experienced users. Complete assembly and cloning protocols are available under Resources at www.idtdna.com/gblocks.

References

  1. Yang L, Yang JL, et al. (2014) CRISPR/Cas9-Directed Genome Editing of Cultured Cells. Curr Protoc Mol Biol, 107(31.1):1–17.
  2. Gibson DG, Young L, et al. (2009) Enzymatic assembly of DNA molecules up to several hundred kilobases. Nature Methods, 6(5):343–345.
  3. Marbiah MM, Harvey A, et al. (2014) Identification of a gene regulatory network associated with prion replication. EMBO J, 33(14):1527–1547.

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