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Although recombinant DNA cloning technology is a core foundational technique in molecular biology, it faces many challenges during actual implementation. Issues such as gene toxicity, insert size limitations, vector capacity bottlenecks, DNA element instability, and complex secondary structures make cloning projects complex and time-consuming. Creative Biogene, with years of accumulated experience and advanced technology platforms, specializes in providing efficient and professional customized gene cloning solutions for clients.
Our professional cloning team can perform customized cloning and subcloning services in client-specified vectors starting from RNA, DNA, or gene sequences. Whether it's simple 125 bp constructs or complex large fragment constructs up to 2 Mbp, we possess the professional technical capabilities to ensure delivery of error-free cloning products. We have completed challenging large-scale complex synthesis projects, including 1.1 Mbp mycoplasma genome creation and stable production of synthetic vaccine viruses, accumulating rich experience in handling complex sequences such as low GC content, high GC content, tandem repeats, palindromic sequences, and homopolymers.
1 PCR Product Cloning Service
Utilizing PCR technology to amplify specific DNA fragments, or performing gene amplification through RT-PCR, followed by cloning target fragments into standard vectors or client-specified vectors. This service is particularly suitable for research projects requiring rapid acquisition of specific gene fragments. Our technical team can optimize PCR conditions according to different experimental requirements, ensuring amplification efficiency and specificity.
2 Vector Insert Fragment Subcloning Service
Professional completion of DNA fragment subcloning operations into target vectors, widely applied in hybrid gene construction and in vivo protein expression systems. Through precise molecular manipulation techniques, we can achieve fragment transfer between different vectors, laying the foundation for subsequent protein expression, functional studies, and other experiments. During subcloning, we strictly control operational conditions to ensure the integrity of the insert fragment and its correct orientation.
3 TA Cloning Technology Service
TA cloning technology takes full advantage of Taq polymerase's terminal transferase activity while lacking 3'-5' exonuclease proofreading activity, enabling the addition of non-template-dependent adenine bases at the 3' end of PCR products. The greatest advantage of this technology is that it requires no primers containing restriction enzyme recognition sequences, no blunt-end treatment of PCR products or adapter addition, and allows direct cloning operations, making it the most convenient and rapid method for cloning PCR products currently available.
4 Synthetic Gene Cloning Service
De novo synthesis of complete genes or DNA fragments and cloning into standard vectors or client-selected vectors, including various expression vectors. This service is particularly suitable for research projects requiring codon optimization, sequence modification, or obtaining naturally non-existent sequences. Our gene synthesis platform can handle various complex sequences, ensuring the accuracy and functional integrity of synthetic products.
Creative Biogene provides an extremely rich selection of cloning vectors, possessing over 100 validated, ready-to-use mammalian expression vectors. The diversity of these vectors stems from our extensive component library, providing unparalleled flexibility and options for your research:
Rich Fluorescent Tag Library
Including GFP, RFP, YFP, BFP, Far-red, Cyan, and other mainstream fluorescent proteins, meeting needs for multi-color labeling, live cell imaging, and deep tissue imaging.
Comprehensive Epitope Tag Library
Providing Flag, His, HA, GST, Myc, Fc, V5, S tags, and other highly efficient options for convenient protein detection, purification, and functional studies.
Diverse Selection Marker Library
Equipped with puromycin, neomycin/G418, blasticidin, hygromycin, and other efficient resistance genes, ensuring rapid and reliable screening of mammalian cell stable lines.
Flexible Inducible/Regulatory Systems
Integrating mature systems Cre-loxP for precise temporal and spatial control of target gene expression.
Extensive Promoter Selection
Including potent CMV, stable EF1α, and PGK, SV40, and other promoters, adapting to different cell types and expression strength requirements.
After clients submit project requirements, our technical expert team conducts a detailed project evaluation, including target gene characteristic analysis, vector selection recommendations, and expression system recommendations. Based on project complexity and client time requirements, we develop optimal technical solutions and project timelines.
Based on requirements analysis results, our molecular biology experts design detailed cloning strategies, including primer design, vector modification, and cloning route planning. For complex projects, we provide multiple alternative solutions to ensure project success rates.
Once projects enter the experimental execution phase, we implement strict quality control and progress monitoring. Each critical milestone has corresponding quality checks to ensure experiments proceed smoothly according to plan. When technical difficulties arise, we promptly adjust strategies to ensure projects are completed on time.
All cloning products undergo strict quality control procedures before delivery, including restriction enzyme digestion verification and Sanger sequencing confirmation. For complex constructs, we also provide NGS sequencing or third-generation sequencing and other high-precision verification services.
Upon product delivery, we provide detailed technical reports including construction strategies, quality control data, and usage recommendations. After delivery, we provide ongoing technical support to assist clients in achieving optimal results in subsequent experiments.
Raw Material Quality Control Standards
| Material Type | Vector Backbone | Vector Components |
| VectorBuilder Stock Materials | Restriction enzyme digestion verification Full-length Sanger sequencing | Full-length Sanger sequencing |
| Client-Provided Materials | Restriction enzyme digestion verification Sanger sequencing (optional) | Full-length Sanger sequencing |
| De novo Synthesized Materials | - | Full-length Sanger sequencing |
Final Vector Quality Control Standards
Thank you for choosing Creative Biogene's gene cloning services. For any related questions or technical consultation needs, please contact our professional technical team. We will wholeheartedly provide you with the highest quality service and most professional technical support. We promise to assist your research projects in achieving success with the highest quality standards, most reasonable prices, and fastest delivery speeds.
Q: Why is my plasmid preparation yield low?
A: Low plasmid yield may be caused by several factors:
1. Origin of Replication Type: The origin of replication on the plasmid determines the plasmid copy number in each bacterial cell. Some origins are inherently low-copy. Please check the copy number characteristics of your plasmid's origin of replication. For low-copy plasmids, increase the E. coli culture volume to obtain satisfactory DNA yields.
2. Insufficient Culture Volume: Please confirm your plasmid preparation column binding capacity and whether the plasmid is high-copy or low-copy. For minipreps, we recommend collecting 1-5 mL of overnight culture. For maxipreps, high-copy plasmids should use 100-150ml overnight culture, while low-copy plasmids should use 300-500ml overnight culture.
3. Antibiotic Degradation: Some antibiotics (particularly ampicillin) degrade rapidly in liquid culture, leading to massive proliferation of plasmid-free bacteria. Please ensure the use of freshly prepared antibiotic-containing media and provide sufficient antibiotic concentration.
Q: Do you provide technical support after delivery?
A: We offer comprehensive post-sale technical support to ensure the success of your experiments, including detailed guidance on vector usage, transformation, and amplification recommendations, expression condition optimization, and troubleshooting assistance tailored to your specific needs.
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