|
Crude Desalted |
RPC Purified** |
Gel Purified |
20 mer oligo
Typical Yield |
30 mer oligo
Typical Yield |
50 mer oligo
Typical Yield |
Scale |
A260 Units |
nmols |
Mg |
A260 Units |
nmols |
mg |
A260 Units |
nmols |
Mg |
50 nmol |
8-10 |
30+ |
0.2-0.3 |
4-5 |
12+ |
0.1-0.16 |
NR* [1-2] |
NR* [2-4] |
NR* [0.03-0.06] |
200 nmol |
20-25 |
80+ |
0.6-0.8 |
8-12 |
24+ |
0.26-0.4 |
4-6 |
8+ |
0.13-0.2 |
1 µmol |
100-120 |
400+ |
3-4 |
40-50 |
90+ |
1.3-1.6 |
20-25 |
40+ |
0.6-0.8 |
Purity & Yield |
| |
Purity is greater than 80%
depending on oligo sequence and structure. Refer to coupling efficiency table for oligo length dependent purity and yield.
No further purification required for PCR and sequencing applications.
Gel purification recommended for oligos above 50 mer and all applications involving cloning and mutagenesis.
**RPC is reverse phase purification using a cartridge; a substitute for HPLC. |
|
|
| |
Purity 85% to 95%
depending on oligo sequence and structure
Yield and Purity will be lower for sequences with high GC content
Not recommended for oligos longer than 35 mer.
**RPC is reverse phase purification using a cartridge; a substitute for HPLC. |
|
|
| |
Purity 98% to ~100%
depending on oligo sequence and structure
Yield will gradually decrease as length of oligo increases. Palindromes, hairpins and high GC content oligos and oligos containing stretches of 3 or more G's induce strong secondary structure and base stacking thus decreasing purity and yield.
*NR Not Recommended |
|
|
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| |
Oligo Size and Purification Recommendations |
Length |
Page |
HPLC/RPC |
8-40 mer |
Yes |
Yes |
41-250 mer |
Yes |
No |
All Gene Link oligos shorter than 40 mer usually do not require any further purification if the application is for PCR or sequencing. |
|
Application Based Purification Recommendations |
Application |
Purification |
PCR and Sequencing |
Not Required |
Cloning and Gene Construction |
Yes |
Mutagenesis |
Yes |
Modified Oligos |
Yes |
Probes |
Yes |
|
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Genelink offers many modifications in oligos. For more details please inquire
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Antisense oligonucleotides refer to short, synthetic oligonucleotide that are complementary in sequence and upon specific hybridization to its cognate gene product induces inhibition of gene expression. Oligonucleotides, as short as 15 mer have the required specificity to inhibit gene expression of a particular gene by annealing to the cellular mRNA (1,2). The mechanism of gene expression is based on two properties; the first is the physical blocking of the translation process by the presence of the short double stranded region, secondly the presence of the RNA-DNA duplex is susceptible to cellular RNase H activity. RNase H cleaves the RNA-DNA duplex region of the mRNA thus preventing the faithful translation of the mRNA (3).
At Gene Link in addition to the synthesis of these modified oligos, we routinely assist customers in the design of the oligos that are particularly suited to their application.
Phosphorothioate Oligos
Propyne dC and dU labeled Oligos
2’O methyl Oligos
Locked Nucleic Acids (LNA) Oligos
2’-5’ linked Oligos
Chimeric Oligos
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The term aptamer is derived from the Latin ‘aptus’ meaning “to fit” and is based on the strong binding of single stranded oligos to specific targets based on structural conformation. Aptamers are single-stranded RNA or DNA oligonucleotides 15 to 60 base in length that bind with high affinity to specific molecular targets; most aptamers to proteins bind with Kds (equilibrium constant) in the range of 1 pM to 1 nM similar to monoclonal antibodies. These nucleic acid ligands bind to nucleic acid, proteins, small organic compounds, and even entire organisms. Aptamers have many potential uses in intracellular processes studies, medicine and technology (1-7, excellent background and review articles).
For your specific APTAMERS please inquire..
For more details please inquire
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