DNA cloning & targeting vector construction
Whether it is direct cloning of large genomic sequences for bioprospecting, chromosomal engineering for industrial strain construction, building complex gene circuits in synthetic biology or assembling CRISPR donor vectors for custom model production, NovoHelix has over 20-years of experience in advanced DNA cloning methodologies. Our deep expertise with genetic tools such as Gibson & GoldenGate assembly, Lambda-red recombineering, linear-linear homologous recombination using modified RecE, or exonuclease-assisted in vitro assembly with recombineering affords us the unique ability to clone and modify large DNA fragments and build complex genetic circuits without scars that would be impossible with traditional cut-and-paste restriction enzyme engineering workflows. Our portfolio also includes chemical gene and fragment synthesis. All constructs are validated by a combination of methods such as Sanger sequencing and restriction endonuclease fingerprinting. Ask us for a custom quote to design and build your next plasmid vector. NovoHelix can also shuttle your vector into a strain of your choice including E. coli BL21 expression strains and the fast-growing bacterium Vibrio natriegens: please see our transformation services.
plasmid DNA transformation
DNA (1µg) with a concentration of at least 20 ng/µl is transformed into the DH5alpha strain of E.coli.
NovoHelix has expertise with chemical transformation, electroporation and transformation by Yoshida effect, i.e. acicular material plus friction. Biolistic transformation service is also possible for some bacterial strains but could incur additional fees.
NovoHelix can transform BACs, linear DNA plasmids, plasmids containing R6K-origin of replication but additional fees may apply. $50 per transformation for standard E. coli strains.
DNA sequencing and analysis ($12/trace)
1. Sanger dideoxy sequencing reactions are performed using the client's template and primer.
- 100-200 ng per reaction of template to be sequenced (PCR product or plasmid)
- sequencing primer - Please note: DNA sequencing primers can be synthesized by NovoHelix at additional cost
2. Analysis of the sequence is performed.
3. Areas of sequence ambiguity will be highlighted.
gene synthesis, size 500 - 5000 bp|$0.55/bp
Plasmid DNA from a miniprep containing the sequence-verified synthesized DNA in the standard cloning plasmid, or an alternate plasmid specified and provided by the client.
- DNA sequence to be synthesized as-is (if no optimization performed).
- Protein sequence to be synthesized.
Please note any flanking DNA sequences to be included, species for codon optimization, and whether specific restriction enzyme sites should be avoided.
Note: Subcloning into a non-standard vector may result in additional costs
Note: Sequence complexity (GC-rich or repeated motifs) may result in additional costs. DNA sequencing and analysis are charged separately.
vector construction (CRISPR donor, DNA ministrings, gene targeting, transgenes)
Inquire about our advanced genetic tools for complex vector construction and these constructs are Sanger or Minion-sequence verified. NovoHelix will provide an in silico map and provide restriction fingerprinting for plasmid structure via 3 digests. DNA is purified by a propriety AEX (anion exchange) chromatography and suitable for microinjection into mouse zygotes or nucleofection/electroporation.