Vector Cloning

Name: Vector Cloning
Brand: VectorBuilder

As the world’s leading provider of custom vector cloning services, VectorBuilder can create any custom DNA vector to meet your research needs. Our platform features about 200 optimized vector systems, validated through in vitro and in vivo (when applicable) testing. Outsourcing your cloning projects to VectorBuilder is often more cost-effective than in-house cloning in terms of reagent and labor cost, with the added benefit of saving your time, effort, and the frustration associated with cloning.

In addition to molecular cloning of vectors, we offer several related services including plasmid DNA preparation, virus packaging, library construction, stable cell line engineering, and therapeutic IVT RNA synthesis.

Highlights

Free, highly-intuitive platform for quick and easy design of custom vectors using our vast inventory of functionally validated vector backbones and components.

Modular “Lego-like” production workflow enabling parallel cloning of custom vectors efficiently and reliably, with over 95% of projects completed on time.

Extensive experience in cloning complex and challenging vectors.

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Full ownership of the IP for your custom vectors and free vector storage.

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Service Information

Overview and workflow of custom cloning services

*Price depends on the type of sourcing, backbone, and cloning type.

Price and turnaround Price Match

While >80% of the vector components can be directly sourced from our in-house collections (based on past production data), we can create vectors from scratch (de novo synthesis) or from customer-supplied materials (vector backbone or gene template). Our custom vector cloning workflow includes QC of customer-supplied materials (if applicable), sourcing of vector components and vector backbones, the actual vector cloning, and final vector QC. A brief overview of price and turnaround for our custom cloning services are shown in Table 1 and 2:

Table 1. Simple vector cloning*

Vector Type	Price (USD)	Turnaround
shRNA vector	$149	4-7 days
gRNA vector (single-gRNA)	$149	4-7 days
gRNA vector (dual-gRNA)	$329	6-12 days
Expression vector	$139	3-6 days

Table 2. De novo synthesis

Fragment Length	Price (USD)**	Turnaround
<1.5 kb	$0.12/bp	5-10 days
1.5-3 kb	$0.14/bp	10-15 days
3-5 kb	$0.17/bp	10-20 days
5-8 kb	$0.21/bp	10-20 days

*Simple vector refers to vectors that can be constructed with 1-step cloning using VectorBuilder's popular in-stock backbones and components.

**The de novo gene synthesis fee may be higher when 1) the fragment contains regions that are difficult to synthesize such as high GC content, simple repeats or segmental repeats; 2) fewer than three DNA fragments are synthesized.

How to Order

View finished vector design > Add vector cloning service and related services to your cart > Get quote or purchase now.

Technical Information

Quality control (QC)

All vectors cloned by VectorBuilder come with a 100% sequence accuracy guarantee. The table below lists our QC items for custom vector cloning:

Raw Material
	Vector backbone	Vector components
VectorBuilder's in-stock material	RE digestion Full-length Sanger sequencing
Customer-supplied	RE digestion Sanger sequencing (optional)	Full-length Sanger sequencing
De novo synthesized	-	Full-length Sanger sequencing
Final Vector
RE digestion Sanger sequencing (depending on cloning method) NGS (optional)

Customer-supplied materials

If customer-supplied materials such as backbones or gene templates are needed, please submit your materials information on "Support" > "Materials Submission Form". Please strictly follow our guidelines to set up shipment to avoid any delay or damage of the materials. All customer-supplied materials undergo mandatory QC by VectorBuilder which may incur an additional QC charge starting from $100 per item, depending on the type and usage of item. Please note that production cannot be initiated until customer-supplied materials pass QC.

Resources

FAQ

What methods does VectorBuilder use for its vector cloning services?

At VectorBuilder, a combination of various molecular biology techniques is utilized to generate custom vectors based on customer needs. While expression vectors are typically constructed using Gateway cloning or Gibson assembly, shRNA and gRNA vectors are cloned using a ligation-based approach to provide you with your desired vectors at the lowest prices and fastest turnaround times utilizing our highly optimized high-throughput cloning platform. Additionally, we routinely use a variety of other methods including de novo gene synthesis and Golden Gate cloning as needed, on a project-by-project basis.

Which bacterial strain does VectorBuilder use for cloning its vectors? What bacterial strains can I use for propagating my plasmid obtained from VectorBuilder?

All vectors at VectorBuilder are cloned using our proprietary host strain VB UltraStable^™, which is designed to achieve high transformation efficiency (>1x10⁹ cfu/ug) and for propagating DNA plasmids with unstable elements such as repeated sequences. Therefore, we highly recommend that you use VB UltraStable^™ chemically competent cells for propagating your plasmids obtained from VectorBuilder. However, our vectors are compatible with and can be propagated in other commonly used bacterial hosts strains such as DH5α and Stbl3.

Why is the yield of my plasmid prep so low?

The origin of replication on your plasmid is for low-copy replication

The number of plasmid copies per bacterial cell is determined by the origin of replication on the plasmid. Some origins have inherent low copy number. Check the copy number of the origin of replication on your plasmid. For low-copy plasmids, increase the amount of E. coli culture for plasmid DNA prep in order to obtain satisfying DNA yield.

Check the copy number of the origin of replication of plasmids supplied by VectorBuilder

The volume of bacterial culture is too low for plasmid prep column

Please check the binding capacity of your plasmid prep column and whether your plasmid is high- or low-copy plasmid. For mini preps, we recommend that you harvest 1-5 ml of overnight bacterial culture. For maxi preps, if the plasmid is high-copy plasmid, we recommend using 100-150 ml of overnight bacterial culture; if the plasmid is low-copy plasmid, we recommend using 300-500 ml of overnight bacterial culture. Typically, for high-copy plasmid, ~5 ug of plasmid DNA can be extracted from every 1 ml of culture in mini prep and ~500 ug of plasmid DNA can be obtained from 150 ml of culture; for low-copy plasmid (e.g. pET), 1.5-2.5 ug of plasmid DNA can be harvested from every 1 ml of culture in mini prep and 150-200 ug of plasmid DNA can be obtained from 150 ml of culture.

Only a fraction of bacteria in the liquid culture contain plasmids

Some antibiotics, ampicillin in particular, degrade fast in liquid culture. As a result, bacteria that do not contain plasmids can propagate to a significant fraction of the culture, causing poor yield of the plasmid prep. To avoid this, please prepare ampicillin containing growth medium freshly before use and make sure that enough ampicillin is supplied. Also, when culturing ampicillin-resistant bacteria, do not let the liquid culture saturate for too long before harvesting. Besides insufficient antibiotics in the culture, extracting plasmid DNA from very old culture can also result in low yield, since many bacterial cells are dead and plasmid DNA they contain is degraded. Therefore, try to extract plasmid DNA from fresh culture. If plasmid prep cannot be performed immediately, you can spin down the bacteria and store the pellet in -80°C freezer for later plasmid prep.

The liquid culture is directly inoculated from E. coli stab culture

Direct inoculation of a liquid culture from the E. coli liquid stock or stab culture you have received from VectorBuilder can very occasionally result in low yield. We recommend streaking the stock onto an LB agar plate containing the appropriate antibiotic first, and then inoculating a liquid culture with a fresh colony growing from that plate. Detailed user instructions can be found by going to menu item Learning Center > Documentation > Stab Culture.

You have not carefully followed the manual of the plasmid prep kit

If you use a plasmid prep kit, please carefully read the manual before use. Improper operations can often lead to poor performance of the kit.

The mini/maxi prep column is low-quality

Some brands of plasmid DNA prep columns perform poorly or inconsistently for DNA preparation.

What happens if there is a mutation in the customer supplied material?

If our QC assays uncover a synonymous mutation, we will contact you for approval in moving forward with cloning, as this mutation will be unlikely to affect experimental outcome. However, if a non-synonymous mutation is found, we will discuss with you the best course of action for correction before cloning and downstream services are initiated.