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Oligonukleotid szintézis, szintetikus biológia

gBlocks® Gene Fragments

Better, faster, more affordable synthetic biology

 

gBlocks® Gene Fragments are double-stranded, sequence-verified genomic blocks that ship in only a few working days for affordable and easy gene construction or modification, applications such as antibody research and CRISPR-mediated genome editing, use as qPCR standards, and more. New! gBlocks Gene Fragments are now available in plates.

 

gBlocks® Gene Fragments Libraries are pools of gBlocks Gene Fragments that contain up to 18 consecutive variable bases (N or K) for recombinant antibody generation or protein engineering. Sequence information is always secure and confidential at IDT. Non-disclosure agreements are available through IDT legal services upon request. twice by UV spectrophotometry to provide an accurate measure of yield. Standard oligos are assessed by mass spectrometry for quality control (QC).

 

gBlocks Gene Fragments are double-stranded DNA molecules of 125–3000 bp in length. gBlocks Gene Fragments are synthesized using the same industry-leading, high-fidelity synthesis chemistry developed by IDT for our Ultramer® oligonucleotides, and are sequence verified prior to shipping. The high sequence fidelity and rapid delivery time make gBlocks Gene Fragments ideal for a range of synthetic biology applications, including the ability to easily assemble multiple gene fragments to reliably generate larger gene constructs.

 

  • Highly versatile - gBlocks Gene Fragments 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 business days (up to 1kb) or 8 business days (up to 2kb).

 

Synthetic Biology Partners

 

biolabsAssembly and cloning of multiple gBlocks Gene Fragments is quick and easy with New England Biolab’s Gibson Assembly® Master Mix

 

syntIDT and SGI are working together to construct custom, synthetic, double-stranded nucleic acids up to 5 kb. IDT uses its custom oligonucleotide manufacturing expertise, including its recently launched gBlocks Gene Fragments product line, in combination with SGI’s proprietary technologies to efficiently assemble small gene constructs.

 

gBlocks® Gene Fragments Libraries

 

gBlocks Gene Fragments Libraries are pools of gBlocks Gene Fragments from 251 to 500 bp in length that contain up to 18 consecutive variable bases (N or K). They are ideal tools for demanding applications such as generating recombinant antibodies or protein engineering.

Below is a representation of the general ordering
format for gBlocks Gene Fragments Libraries:

nnn

Need something else? Tell us about it! 

We realize that this first offering for gBlocks Gene Fragments libraries is somewhat limited. We are working hard at offering more complex libraries in the future. Help us prioritize by sharing what you need from us in as much detail as possible (i.e., show us sequences including mixed bases, annotations, drawings, etc.) and send to .

 
Ordering format for gBlocks Gene Fragments Libraries

gBlocks Gene Fragments Libraries are entered the same way as gBlocks Gene Fragments. The ordering interface now accepts N or K (G or T) mixed bases when present in an acceptable format (K mixed bases placed in the third position of codons eliminate two of the three stop codons from being included in the gene fragments libraries). The gBlocks Gene Fragments Libraries can be 251 to 500 bp in total length, including the variable region. The variable regions can be up to 18 consecutive N or K bases long, and needs to be at least 125 bp from either end of the gene fragment.

 

Why did IDT 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 418gene 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.

 

How to use gBlocks Gene Fragments Libraries

gBlocks Gene Fragments Libraries can be used directly in some suitable applications such as In vitrotranscription or when used as standard or internal controls for qPCR or NGS experiments. More often, they are assembled into functional genes or longer constructs by seamless assembly techniques or traditional restriction cloning.

 

Adding diversity at reasonable cost

IFor 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.100 nmole and 250 nmole scales and up to 40 residues on the 1 µmole scale, with a maximum of 4 unique Hand Mixes per oligonucleotide.When entering your sequence, use the “Mixed Base” tab at the bottom of the page. It lists the IUPAC-IUB symbols and is where custom mix ratios should be entered.

 

Quality Assurance

Each gBlocks Gene Fragment ordered from IDT will go through the following verification process:

  • Researchers enter the gene fragments sequences (125–3000 base pairs) 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 not currently available; no “modified" bases (Inosine, Uridine).
  • Automated review by screening tools at the time of order entry and, and expert review by IDT scientists, of entered sequences for characteristics that may interfere with synthesis
  • 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.

 

QC procedure and sequence verification

Each gBlocks Gene Fragment undergoes the following quality control procedures:

  • Size verification by capillary electrophoresis
  • Sequence identification by mass spectrometry
  • Final consensus sequence verification

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 have reduced fidelity, which can be addressed by the end user sequencing additional clones.

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

 

Guaranteed yield

  • gBlock Gene Fragments are delivered dry, normalized to 250, 500, or 1000 ng, depending on length.
  • gBlock Gene Fragments Libraries are delivered dry, normalized to 200 ng.
  • gBlocks Gene Fragments in Plates are delivered in 25 µL of nuclease free water, normalized to 10 ng/µL.

 

Confidentiality

  • All sequence information is kept confidential at IDT.
  • All online ordering steps, including sequence entry and choice of parameters, are also secure and protected.

 

Possible applications of gBlocks Gene Fragments include, but are not limited to:

  • Protein Expression
    • Recombinant antibodies
    • Novel fusion proteins
    • Codon optimized short proteins
    • Functional peptides: catalytic, regulatory, binding domains
  • microRNA genes
  • Template for in vitro transcription (IVT)
  • Regulatory sequence cassettes
  • Microarray-ready cDNA
  • Gene variants and SNPs
  • DNA vaccines
  • Standards for quantitative PCR and other assays
  • Functional Genomics
    • Limitless flexibility for protein mutagenesis
      • Mutant libraries
      • Deletion mutants

 

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

 

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