Home ProductsOligo SynthesisProportional Control Degenerate Primers DNA Oligo Pool Synthesis Oligo Synthesis
I'm Online Chat Now

Proportional Control Degenerate Primers DNA Oligo Pool Synthesis Oligo Synthesis

Proportional Control Degenerate Primers DNA Oligo Pool Synthesis Oligo Synthesis
Proportional Control Degenerate Primers DNA Oligo Pool Synthesis Oligo Synthesis

Large Image :  Proportional Control Degenerate Primers DNA Oligo Pool Synthesis Oligo Synthesis

Product Details:

Brand Name: CELLFREE
Model Number: CF-OL-004

Payment & Shipping Terms:

Packaging Details: vial
Delivery Time: 10 business days
Detailed Product Description
Material: Primers Appearance: Lyophilized Powder
Package: Tube QC: COA
• Standard Service: Free Design Super Service: Proportional Control
High Light:

Degenerate Primers DNA Oligo Synthesis

,

Degenerate Oligo Pool DNA Synthesis

,

Proportional Control Oligo Synthesis

Degenerate Primers Custom Degenerate Oligo Pool Synthesis DNA Oligo Synthesis

DNA oligos are chemically synthesized with four deoxyribonucleotides of A, T, C and G. These sequences are widely used in various ways in field of biology. CELLFREE is confident in our ability to provide high-quality DNA oligo products to scientific researchers and industrial customers around the world. With the combination of our state-of-the-art synthesizer, advanced production technology and professional technical support team we have consistently achieved low mutation rates, short turnaround times, high cost performance.

 

Genetic codes have the tendency to degenerate, which lead to the use of degenerate primers in PCR experiments to exhibit specific genes. CELLFREE provides both the design and synthesis of these degenerate primers, as well as custom degenerate oligo pool synthesis upon request.

 

Introduction

 

A degenerate primer is defined as: “A mix of oligonucleotide sequences in which some positions contain a number of possible bases, giving a population of primers with similar sequences that cover all possible nucleotide combinations for a given protein sequence” (Iserte 2013). For example:

ATCGTT[GC]AAGT[AGC]ATC

refers to a series of primers in which the seventh and twelfth nucleotides are degenerate. The amount of degeneracy is defined by the number of different primer combinations in the mix. The above example has a degeneracy value of six.

 

Service list

  • Standard Service: No design fee, suitable for more than 95% PCR projects.
  • Superior Service: Utilized for certain projects sensitive to codon bias. CELLFREE can significantly reduce the deviation inherent in the synthesis procedures and realize the accurate proportional control.
Symbol Base Type Standard Ratio Price
Standard Level Superior Level
R A,G 50% : 50% Free Ask for quotation
Y C,T 50% : 50%
M A,C 50% : 50%
K G,T 50% : 50%
S C,G 50% : 50%
W A,T 50% : 50%
H A,C,T 33% : 33% : 33%
B C,G,T 33% : 33% : 33%
V A,C,G 33% : 33% : 33%
D A,G,T 33% : 33% : 33%
N A,C,G,T 25% : 25% : 25% : 25%

* Non-standard base proportion should be declared clearly, for example, N(20%A:30%C:40%G:10%T).

 

Designing degenerate primers

 

1) Align multiple amino acid sequences using free online software.
2) Target an area approximately 200-500 base pairs in length for optimal PCR amplification.
3) Position forward and reverse primers in more conserved regions – the less degenerate, the further apart these can be.
4) Include between 6 and 7 amino acids in the primers, equating to ~15-20 base pairs.
5) Try to include amino acids methionine and tryptophan, which are coded by a single codon (three-letter nucleotide code), and avoid amino acids leucine, serine and arginine, which can each be coded by six codon combinations.
6) For subsequent cloning procedures and to increase primer length (and therefore annealing temperature) add a 5’ tail (6-9 base pairs) containing a restriction enzyme site.
7) If there is complete degeneracy (no matches among any given species), consider using the base inosine (structurally similar to guanine) as it can pair with any of the four bases, although it will bind to cytosine preferentially. Alternatively, insert N for aNy base to ensure equimolar concentrations of each base at that position in your primer mix.
8) Avoid degeneracy at the 3’ terminus (this would not be a good place to insert inosine).

Contact Details
CELLFREE SCIENCE CO., LTD.

Contact Person: Pan Lin

Send your inquiry directly to us (0 / 3000)