Gateway Cloning Service is a specialized service offered by our company that allows for efficient and precise transfer of DNA fragments into a variety of expression vectors. This technique is widely used in molecular biology research and enables seamless assembly of DNA constructs. Whether you need to clone small or large DNA fragments, our Gateway Cloning Service provides a reliable and versatile solution.
Service Process
Our Gateway Cloning Service follows a systematic process to ensure a seamless and efficient cloning experience. Here is an overview of the steps involved:
- Consultation: We start by engaging in a detailed consultation to understand your project requirements and goals. This allows us to provide personalized recommendations and insights tailored to your research needs.
- Design and Primer Synthesis: Our experts utilize their expertise to design the appropriate Gateway Cloning strategy for your project. We also synthesize the required primers necessary for the cloning reaction.
- Cloning Reaction: This is where the magic happens. We perform the Gateway Cloning reaction, transferring your DNA fragment of interest into the desired vector. Our experienced scientists ensure accurate and precise fragment insertion.
- Transformation and Selection: To ensure successful cloning, we transform the recombinant vectors into competent cells. These cells are carefully selected to ensure the stable integration of your DNA construct.
- Confirmation and Analysis: We understand the importance of verifying the correct insertion of your DNA fragment. Our team employs various molecular biology techniques, such as restriction digestion and sequencing, to confirm the integrity and accuracy of your cloned vector.
- Deliverables: Once the cloning process is successfully completed, we provide you with the cloned vectors along with detailed documentation of the cloning process. This documentation serves as a valuable resource for your research and experimental records.
For more information about our Gateway Cloning Service and to discuss your project requirements, please contact our team by submitting your contacts, We are here to assist you every step of the way.
Case Studies
Multisite Gateway-mediated Cas9/gRNA expression system (GCas method). (A) sgRNA module vectors harboring the att sites. U6, mammalian U6 type III RNA polymerase III promoter; H1, H1 RNA polymerase III promoter; BbsI, a single sgRNA cloning site mediated by BbsI digestion; BsmBI, a dual sgRNA cloning site mediated by BsmBI digestion; Scaff, scaffold sequence of sgRNA followed by TTTTTT (polyT terminal signal of RNA polymerase III); C4s, a Csy4-mediated RNA cleavage site. See Figure E in S1 Supporting Information and the corresponding legend for the cloning of sgRNA(s) into the sgRNA module vectors. (B) Promoter vectors harboring attL5 and attR3. CMV, cytomegalovirus promoter; CAG, cytomegalovirus enhancer fused to the chicken beta-actin promoter and introns of chicken beta-actin and rabbit beta-globin; EF1α, Elongation factor 1-alpha promoter. (C−D) Vectors harboring Cas9 alone or Cas9/EGFP harboring attL3 and attL2. 2As are illustrated as right-pointing triangles. Cas9, spCas9; 2A, Thosea asigna virus 2A peptide sequence (T2A); Csy4, Csy4 RNase of Pseudomonas aeruginosa; FokI, dimerization-dependent wild-type FokI nuclease-domain; Cas9n, mutated Cas9 nickase (D10A); dCas9, catalytically inactive Cas9. (E) Destination vectors for the GCas method. The CmR and ccdB gene are located between attR1 and attR2 in each destination vector (not described) for the positive/negative selection of transformants harboring the vector itself or the desired Cas9/gRNA expression vector. Gray arrows and boxes in CSIV-DEST indicate regulatory sequences for lentivirus production such as 5'LTR (5'-side long terminal repeat), C (Packaging signal), cPPT (central polypurine tract), WPRE (woodchuck hepatitis virus posttranscriptional regulatory element) and 3'LTR (3'-side long terminal repeat). 5'ITR and 3'ITR, 5'-side and 3'-side of Piggybac inverted terminal repeats. The entire sequences of the vectors described are attached in the S1 sequence. (F) Other module vectors for the GCas method. pA, simian virus 40 polyadenylation signal sequence.——(S Yoshimatsu,2019)
FAQs
Here are some frequently asked questions about our Gateway Cloning Service:
What types of vectors can be used in Gateway Cloning?
Gateway Cloning is compatible with a wide range of expression vectors, including plasmids and viral vectors. Our experts can guide you in selecting the most suitable vector for your specific application.
What is the turnaround time for Gateway Cloning Service?
The turnaround time for Gateway Cloning can vary depending on the complexity of the project. During the consultation phase, our team will provide you with an estimated timeline based on the specific requirements of your project.
Can you clone large DNA fragments using Gateway Cloning?
Absolutely! Gateway Cloning is suitable for cloning both small and large DNA fragments. Our experts have the expertise and resources to handle a wide range of fragment sizes.
Do you offer downstream services, such as protein expression and purification?
Yes, we provide comprehensive services for downstream applications. In addition to Gateway Cloning, we offer services such as protein expression and purification. Our goal is to provide a holistic solution to meet your research needs.
How do I get started with Gateway Cloning Service?
Getting started with our Gateway Cloning Service is simple. Just contact our team to discuss your project requirements, and we will guide you through the process. Our experts are here to assist you every step of the way.
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Schematic diagram describing the Gateway cloning
