NEB® PCR Cloning Kit (Without Competent Cells)
Product informationCode | Name | Size | Quantity | Price | |
---|---|---|---|---|---|
E1203S |
NEB PCR Cloning Kit (Without Competent Cells) |
20 rxns | - | Unavailable in your region |
NEB® PCR Cloning Kit (Without Competent Cells)
Allows for in vitro transcription. Also available with competent cells.
Product Introduction
Easy cloning of all PCR products, including blunt and TA ends
- Updated to allow for in vitro transcription with both SP6 and T7 promoters
- BsaI-site removed from the ampicillin-resistant gene (allows for the cloning of Golden Gate Assembly modules)
- Fast cloning experiments with 5-minute ligation step
- Simplified screening with low/no colony background and no blue/white selection required
- Save time by eliminating purification steps
- Flanking restriction sites available for easy subcloning, including choice of two single digest options
- Provided analysis primers allow for downstream colony PCR screening or sequencing
- Ready-to-use kit components include 1 kb control amplicon, linearized cloning vector and single-use competent E. coli
- Longer shelf life (12 months), as compared to some commercially available products
- Also available with competent cells
Catalog # | Size | Concentration |
---|---|---|
E1203S | 20.0 reactions |
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How Does the NEB® PCR Cloning Kit Work?
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NEB® TV Ep. 8 – Cloning and DNA Assembly
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Overview of PCR Cloning
- Product Information
- Protocols, Manuals & Usage
- FAQs & Troubleshooting
- Citations & Technical Literature
- Quality, Safety & Legal
- Other Products You May Be Interested In
Product Information
Description
- Updated to allow for in vitro transcription with both SP6 and T7 promoters
- BsaI-site removed from ampicillin-resistant gene (allows for cloning of Golden Gate Assembly modules)
- More restriction sites added, including four 8-base cutters
The ultimate in flexibility: clone with any amplicon made with any DNA polymerase, with or without 5´ phosphates, purified or not!
- This product is related to the following categories:
Kit Components
Kit Components
The following reagents are supplied with this product:
Properties & Usage
Features
- In vitro transcription with both SP6 and T7 promoters
- BsaI-site removed from the ampicillin-resistant gene (allows for the cloning of Golden Gate Assembly modules)
- Easy cloning of all PCR products, including blunt and TA ends
- Fast cloning experiments with 5-minute ligation step
- Simplified screening with low/no colony background and no blue/white selection required
- Save time by eliminating purification steps
- Flanking restriction sites available for easy subcloning, including choice of two single digest options
- Provided analysis primers allow for downstream colony PCR screening or sequencing
- Ready-to-use kit components include 1 kb control amplicon, linearized cloning vector and single-use competent E. coli
- Longer shelf life (12 months), as compared to some commercially available products
Related Products
Companion Products
- Quick-Load® Purple 1 Kb Plus DNA Ladder
- LongAmp® Taq 2X Master Mix
- OneTaq® 2X Master Mix with Standard Buffer
- LongAmp® Taq DNA Polymerase
- OneTaq® DNA Polymerase
- Deoxynucleotide (dNTP) Solution Mix
- NEB® 10-beta Competent E. coli (High Efficiency)
- HiScribe® T7 ARCA mRNA Kit (with tailing)
- HiScribe® T7 ARCA mRNA Kit
- HiScribe® T7 Quick High Yield RNA Synthesis Kit
- HiScribe® T7 High Yield RNA Synthesis Kit
- SP6 RNA Polymerase
- T7 RNA Polymerase
- Ribonucleotide Solution Mix
- RNase Inhibitor, Murine
- RNase Inhibitor, Human Placenta
Product Notes
- The NEB PCR Cloning Kit contains a sufficient supply of materials to perform 20 x 10 μl cloning reactions. Primers are also provided, allowing screening for inserts by colony PCR and/or sequencing.
References
- Wang, Y. et al. (2004). Nucleic Acids Research. 32, 1197-1207.
- Heurgue-Hamard, V. et al. (2000). The EMBO Journal. 19, 2701-2709.
- Tenson, T. et al. (1999). Journal of Bacteriology. 181, 1617-1622.
Protocols, Manuals & Usage
Protocols
Manuals
The Product Manual includes details for how to use the product, as well as details of its formulation and quality controls.
Usage & Guidelines
FAQs & Troubleshooting
FAQs
- How does the NEB® PCR Cloning Kit work?
- How can the cloning vector work with both blunt-ended amplicons and single-base overhang-containing amplicons?
- Do these polishing components present in the master mix affect my cloning efficiency if my insert already has blunt ends?
- Do my inserts have to possess 5´ phosphates?
- Can the cloning kit be used for inserts that are not necessarily PCR amplicons?
- Can the cloning kit be used with inserts containing 5´ or 3´ overhangs greater than the single-base overhang achieved by PCR with Taq DNA polymerase?
- Can I use the NEB PCR Cloning Kit featuring pMiniT 2.0 for Golden Gate Assembly?
- Does the PCR product need to be purified?
- Can I use a different competent E. coli strain than the provided NEB 10-beta strain?
- How can I maximize the number of transformants?
- Can I scale down the reactions to use less vector?
- Are there limits regarding the size of inserts that can be cloned?
- Are the NEB 10-beta Competent E. coli (Cloning Efficiency) provided in the kit the same cells as NEB 10-beta Competent E. coli (High Efficiency)?
- How can I determine if my NEB 10-beta cells are competent?
- Can Cloning Mix 1 and Cloning Mix 2 be mixed together before adding them to the ligation reaction?
- Where are the +1 transcription positions for the SP6 and T7 promoters for in vitro transcription and translation located?
- What is the difference between the original pMiniT and the pMiniT 2.0 linearized vector backbone now provided in the kit?
- When cloning a PCR fragment into the pMiniT 2.0 vector provided with the NEB PCR cloning kit, where in the multiple cloning site (MCS) of the vector is the insert ligated?
Citations & Technical Literature
Citations
Additional Citations
Quality, Safety & Legal
Quality Assurance Statement
Quality Control tests are performed on each new lot of NEB product to meet the specifications designated for it. Specifications and individual lot data from the tests that are performed for this particular product can be found and downloaded on the Product Specification Sheet, Certificate of Analysis, data card or product manual. Further information regarding NEB product quality can be found here.Specifications
The Specification sheet is a document that includes the storage temperature, shelf life and the specifications designated for the product. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]Certificate Of Analysis
The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality controls for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]- E1203S_v1_10009599
- E1203S_v1_10013259
- E1203S_v1_10022017
- E1203S_v1_10031310
- E1203S_v1_10035043
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- E1203S_v1_10164950
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- E1203S_v1_10176797
- E1203S_v1_10182438
- E1203S_v1_10187352
- E1203S_v1_10202790
- E1203S_v1_10207429
- E1203S_v1_10221514
- E1203S_v1_10238527
- E1203S_v1_10245961
- E1203S_v1_10264548
Safety DataSheets
The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.Linearized pMiniT™ 2.0 Vector
Amplicon Cloning Control
Cloning Analysis Reverse Primer
Cloning Analysis Forward Primer
Cloning Mix 1
Cloning Mix 2
Legal and Disclaimers
Products and content are covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB). The use of trademark symbols does not necessarily indicate that the name is trademarked in the country where it is being read; it indicates where the content was originally developed. The use of this product may require the buyer to obtain additional third-party intellectual property rights for certain applications. For more information, please email [email protected].This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.
New England Biolabs (NEB) is committed to practicing ethical science – we believe it is our job as researchers to ask the important questions that when answered will help preserve our quality of life and the world that we live in. However, this research should always be done in safe and ethical manner. Learn more.
Other Products You May Be Interested In
The supporting documents available for this product can be downloaded below.