Primer synthesis is only one step in the process of whole gene synthesis service, which requires splicing, cloning, screening, QC validation and finally delivery of finished plasmids, so the cost of whole gene synthesis is higher.

High GC sequence: usually refers to sequences with GC content >70% or sequences with GC content >80% within 100 bp in a row;
Low GC sequence: usually refers to genes with <25% of full-length GC content or sequences with <15% of GC content within 100 bp in a row;
Long sequence genes or vectors that are too large (>10 kb) resulting in subclonal splicing difficulties;
Special vector elements or special protein genes (e.g. lethal genes, ion channel proteins, gene methylation, repetitive sequences in vectors, etc.);
Repetitive fragment genes: containing forward or reverse repeats;
Complex genes such as genes containing multiple consecutive bases (e.g. Poly A termination signal).

1. Gibson assembly: Based on 5' nucleic acid exonuclease, DNA polymerase and ligase, DNA assembly is realized by using homologous sequences at both ends of DNA fragments (generally 15-40 bp in length);
2. Golden Gate assembly: utilizing type IIS restriction enzymes to cut outside the recognition site, designing specific protruding sequences to assemble multiple fragments at the same time, and enzymatic cleavage and ligation can be carried out at the same time;
3. SLIC: utilizes specific nucleic acid fragments at the ends of DNA to attract each other and joins DNA fragments together by ligase;
4. Depending on the size and complexity of the gene there are also methods such as homotailase BioBrick, UNS, etc.

For the sample plasmid or bacterial liquid demand provided by customers:
1. bacterial solution: please add 200-500ul of overnight culture bacterial solution to sterilized glycerol to a final concentration of 15-20%, and store it in a 1.5 mL or 2 mL centrifuge tube, pay attention to the seal to avoid contamination;
2. Plasmid: 2-5 μg of plasmid solution dissolved in sterile water/low TE buffer or lyophilized plasmid should be provided, pay attention to sealing to avoid contamination;
Please indicate the name of the sample and resistance on the wall of the tube, provide the sequence map of the plasmid as well as the sequence of the vector backbone with not less than 50 bp upstream and downstream of the gene cleavage site, to ensure that the gene is accurately cloned in the specified vector location, and analyze the distribution of the corresponding cleavage site in the target sequence and vector.

1. Full gene synthesis delivery of finished product includes:
1) One tube of lyophilized plasmid DNA (about 2-4 μg);
2) One tube of glycerol bacteria corresponding to plasmid (200-500ul);
(3) One copy of COA of quality inspection report form;
(4) A delivery note;
2. The shipping documents (electronic version) include the standard sequence, sequencing results, sequencing map, plasmid structure map, plasmid full-length DNA file, and sample quality inspection report (COA);
Upon completion of the project, these documents will be packaged and sent to your email address by email.