Gene Synthesis and CDNA(ORF) cloning
Introduction to Gene Synthesis and CDNA(ORF) cloning
Gene synthesis is a synthetic biological technique, which is the most widely used in biological research. The synthesis of DNA encoding regulatory elements, genes, pathways and entire genomes provides us powerful ways for both test biological hypotheses and harness biology. The emerging field of synthetic biology is generating insatiable demands for synthetic genes. Our company offers you high quality gene synthesis service, and can help you from the planning stage to experimentation.
1. Sending gene sequence and requirements to [email protected]
2. A technical expert will analyze your target sequence and provides a detailed proposal, including sequence design, time line and final cost.
3. Your review and approval of the experimental proposal.
4. Production starts immediately, weekly production report.
5. Delivery, you will receive:
a) The overall report of gene synthesis
b) 4 µg of lyophilized plasmid containing your gene insert
c) Sequence chromatograms covering your gene
d) Construct map for the plasmid
|Products delivered by Gene synthesis services|
|Product composition||1.Plasmid: 2-5µg High-purity lyophilized plasmid DNA containing your intended synthetic construct
2.Sequencing results: Electronic version abi, seq file.
Gene Synthesis Report sheet: Electronic PDF file and paper version A4 paper
|Form||4 µg of lyophilized plasmid containing your gene insert;
Sequence chromatograms covering your gene;
Construct map for the plasmid;
|Instructions for use and Storage||1.According to your experiment requirements plasmid is dissolved in ddH2O with the appropriate concentration.
Plasmid can be quantitatively dissolved and preserve at -20℃.
Sequencing results: Use Seqman to compare your target sequence with the sequencing results.
1. Reliable: 1.skillful staffs with more than 10years experiences.
2. Low Cost: competitive 0.2$ /bp for long genes.
3. Guaranteed sequence: 3.cutting-edge gene synthesis platform ensures mutation-free DNA synthesis, for any length and any complexity.
4. Free additional services: vector designs, codon optimizations, gene cloning solutions, anything that helps you to finish experiments.
5. Various vectors: providing vector choices, no more secondary cloning.
6. Easy online ordering: getting a quote now.
Applications and Figures
Frequently Asked Questions(FAQs)
- 1. How does your company synthesize genes?
The project manager will analyze the sequence submitted by the customer, according to the difficulty degree of different sequences, complete primer design, synthesis, primer splicing and PCR amplification, and finally connect the target gene to the corresponding vector, and through the enzyme cutting QC identification, Sanger sequencing verification to ensure the correctness of the sequence.
- 2. How long can your company synthesize a gene?
We have a leading gene synthesis platform with a maximum synthetic gene length of up to 50kb.
- 3. Can your company synthesize some of the more difficult genes, such as high GC, repeat sequence?
Yes. Our company has a gene synthesis platform that has successfully completed the synthesis of difficult genes for many international clients, such as high GC content sequences, low GC content sequences, forward or reverse repeating sequences, etc. At the same time, we will carefully evaluate each customer's difficulty sequence, provide customers with professional technical support and free codon optimization services, and provide detailed (including time and price information) of the quotation, for your project to create a professional experimental program.
- 4. Is there any additional charge in addition to the cost of synthesizing each base pair?
The project manager will provide you with a reasonable quotation based on the sequence analysis results and the gene synthesis fee. Synthesis of conventional genes (no high GC content, no low GC content and no forward or reverse repetition sequence, etc.) and choose our company to provide free carrier to complete the cloning service, our company will be based on the base unit price and serial length as a charge, to give the final price; synthetic difficulty gene (high GC content, Low GC content and forward or reverse repeats, etc.) or the order contains services such as sub-cloning requirements, we will list in the quotation other than the genetic synthesis of fees, and give a final quotation.
- 5. Does your company provide cryptographic sub-optimization services?
Our company has a professional bio-information team and a leading gene synthesis platform, we provide customers with free codon optimization services. The project manager will select the most appropriate optimized host based on the customer's research needs.
- 6. Is it necessary to optimize the codon?
In most cases, it is necessary for genes used in protein expression. The genes of eukaryotic organisms need to be expressed in prokaryotic organisms. Because of the codon preference of eukaryotes is very different from that of prokaryotic organisms, codon optimization of the genes will significantly improve the expression efficiency.
- 7. What information do I need to provide for genetic optimization?
If genetic optimization is needed, we need you to provide the following information: a. Optimized gene sequence or protein sequence; b. optimized host; c. The enzyme-cutting site to be added at both ends of the gene, or the enzyme-cutting site to be removed within the gene; d. Whether a specific termination codon is required; e. If you need to add a Kozak sequence, we generally recommend that you add
- 8. What are the effects of codon optimization on gene expression?
The codon optimization platform has thousands of optimized host and selection parameters, which can be optimized according to the different needs of the customer, choosing different hosts and parameters. The main parameters of the sequence optimization reference are as follows:
1) codon preference;
2) GC content;
3) mRNA level two structure
4) User-defined parameter selection;
5)Repeat sequence (forward repetition, reverse repetition, palindrome sequence);
6)Enzyme cut-off site (delete or insert).
- 9. What kind of termination codon is preferred in E. coli expression system? What’s about the mammalian system?
in E. coli, tag is seldom used, and taa/tga is often used. In mammals, TGA is used more frequently than TAA and tag. Compared to the difference between mammals and E. coli, codon usage tables differ greatly between different organisms in mammals.
- 10. What is the carrier of your company?
Free delivery of pUC57 vector, assembled the whole gene will be cloned to the pUC57 vector, through the enzyme cut for QC identification, Sanger sequencing verification to ensure the correctness of each sequence.
- 11.What is the difference between the two vectors of puc57-simple and Puc 57? Under what circumstances is it advisable to use the Puc57-simple carrier?
In addition to the MCS area, the Puc 57-simple carrier has retained only two common enzyme cutting sites of NdeI and EcoRV in the Puc 57 carrier base. B. Provide free pUC57 standard carrier. In general, the synthesized gene will be cloned into the SMA I or EcoRV sites of the standard carrier. We recommend that customers use the Puc57-simple carrier if they want to avoid some of the commonly used enzyme cut sites on the vectors to facilitate subsequent sub-cloning needs.
- 12. What antibiotic resistance gene is contained on the pUC57 carrier?
- 13. Can I clone the synthesized gene to the vector I specified?
Yes. In addition to pUC57 and puc57-simple, other carriers need to be provided by the customer themselves. If you need to clone a synthesized gene onto a vector you specify, you need to provide information about the carrier, and we will charge the corresponding sub-cloning fee based on the length of the target gene.
- 14. How does your company determine the correctness of the final synthetic gene sequence?
In the entire process of gene synthesis, we take strict and effective quality control to ensure the accuracy of synthetic genes. First, before the project starts, the project manager will analyze your sequence, write the order according to your needs, and then ask you to verify that the sequence information and requirements for the order are correct. When the order is confirmed, our experimenter will complete the sequence splicing, the whole gene cloning to the target vector, and through the enzyme cut for QC identification, Sanger sequencing verification to ensure the correctness of the sequence. Finally, we will send the order delivery document and the delivery form to your email address.
- 15. What method does your company recommend for DNA Resuspension?
Our recommendation is to take the following steps for DNA resuspension:
1)After receiving the sample, before opening the sample cap, centrifuge the sample tube at 12000 rpm for 1 minutes to prevent the loss of freeze-dried plasmid when opening the lid, to ensure that the lyophilized plasmid DNA is located at the bottom of the collecting tube;
2) after centrifugation, add a certain volume of sterilized double steam water or TE buffer to the sample.