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Ph.D.™ Peptide Display Cloning System

Catalog # Concentration Size List Price Quantity Your Price
E8101S 20 µg $318.00
$286.20
Catalog # Size List Price Your Price
E8101S 20 µg $318.00
$286.20
Catalog #
Qty:
 
*On-line ordering is for Canadian customers only. Web pricing is applicable only to orders placed online at www.neb.ca

The Ph.D.™ Peptide Display Cloning System contains M13KE vector, the parent vector of NEB’s Ph.D. libraries. The included M13 extension primer is used to make DNA duplex out of a user provided synthetic oligo, with or without random bases. The display system is best used to display small peptides, i.e. sequences of 50 amino acids or less.

  • M13KE Vector is based on M13mp18 containing the entire M13 genome, with specific cloning sites at the 5’ gIII, encoding pIII coat protein
  • Allows for user-designed custom peptide library construction
  • May be transfected into F’ E. coli to obtain infectious phage
M13KE is a simple M13 derivative in which cloning sites have been introduced at the 5´ end of gene III for display of short peptide sequences as N-terminal pIII fusions. Because this is a phage, rather than a phagemid vector, all 5 copies of pIII on the surface of each virion will be fused to the cloned peptide. Since displayed proteins longer than 20–30 amino acids have a deleterious effect on the infectivity function of pIII, this vector is suitable only for the display of short peptides. Additionally, the vector does not carry a plasmid replicon or antibiotic resistance, so it is necessary to propagate the vector as phage, rather than a plasmid (i.e., titer for plaques, not colonies). This simplifies the intermediate amplification steps during panning considerably, as it is not necessary to express antibiotic genes before plating, or to use helper phage during amplification. The steps necessary to clone a peptide library into M13KE are outlined below. To clone a single peptide sequence, reactions can be scaled down.

Figure 1
figure 1
Construction of a peptide library in M13KE. Schematic shows the sequence of the peptide cloning site as well as the strategy for designing and cloning a peptide library into M13KE. The sequence of the extension primer is outlined. N = A, G, C or T; X = any user defined or randomized amino acid.
Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration
  M13KE gIII Cloning Vector N3541SVIAL -20 1 x 20 µg 1 mg/ml
  M13 Extension Primer S1203AVIAL -20 1 x 2,150 pmol 40 pmol/μl

Properties & Usage



Companion Products
References
  • Sambrook, J. and Russell D.W (2001). Molecular Cloning: A Laboratory Manual, (3rd ed.). Cold Spring Harbor, Cold Spring Harbor Laboratory Press.
Citations
Additional Citations
  • Kaur K, Taneja NK, Dhingra S, Tyagi JS (2014) DevR (DosR) mimetic peptides impair transcriptional regulation and survival of Mycobacterium tuberculosis under hypoxia by inhibiting the autokinase activity of DevS sensor kinase BMC Microbiol 14, 195.PubMedID: 25048654, DOI: 10.1186/1471-2180-14-195
Quality Control Assay
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]
Safety Data Sheets
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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.

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