Purification of oligonucleotides

• Molecular Biology
  
• HPLC
  
• RNA HPLC
  
• DS (desalting)
  
• Cell culture
  
• PAGE
  
• Reverse phase C18
  
• Documentation
  

• Deprotection of the 5'-hydroxy group
• Coupling of the appropriate building block
• Capping of unreacted 5'-hydroxy groups
• Oxidation of the phosphittriester group to phosphatetriester

Molecular biology grade purification

Salts, base protecting groups and short failure sequences up to 6 bases are removed efficiently. The amount of full length product depends on chain length and coupling yield. A typical 20mer with an average coupling yield of 98.5 % is composed to 70 % of full length product. This purification is recommended for oligonucleotides with a chain length up to 30 bases for standard molecular biology applications such as PCR, sequencing, etc.

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product	scale	amounts [oD]*	price/oligonucleotide
Molecular biology grade purification	0.01 µmol	0.5 -1.0	no charge
	0.05 µmol	2.0 -4.0	no charge
	0.20 µmol	10.0 -20.0	no charge
	1.00 µmol	50.0 -100.0	no charge
	15.00 µmol	600.0 -1200.0	no charge

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HPLC grade purification

Apart from salts and base protecting groups, the trityl-on HPLC step efficiently removes trityl-off failure sequences. The purity of the product depends on the nature and length of the sequence. IBA NAPS recommends this purification for all applications where full-length or high reproducibility are essential, including: cloning, PCR, mutagenesis, probes, modified oligonucleotides and oligonucleotides longer than 40 bases. This method is not able to remove trityl-on failure sequences.

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product	scale	amount	cat. no.	price/oligonucleotide
HPLC grade purification	0.01 µmol	1-2 nmol	5-1020-001	9.00 EUR
	0.05 µmol	5-10 nmol	5-1020-002	9.00 EUR
	0.20 µmol	25 -50 nmol	5-1020-003	10.00 EUR
	1.00 µmol	100-200 nmol	5-1020-004	15.00 EUR
	15.00 µmol	1500-3000 nmol	5-1020-005	184.00 EUR

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Double HPLC grade purification

This protocol consists of a trityl-on HPLC purification and a subsequently performed trityl-off HPLC purification. The second step removes efficiently oligonucleotides with base modifications and partly trityl-on failure sequences.

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product	scale	amount	cat. no.	price/oligonucleotide
Double HPLC grade purification	0.01 µmol	0.5-1 nmol	5-1030-001	18.00 EUR
	0.05 µmol	2.5-5 nmol	5-1030-002	20.50 EUR
	0.20 µmol	10.0-25 nmol	5-1030-003	25.00 EUR
	1.00 µmol	50.0-100 nmol	5-1030-004	30.00 EUR
	15.00 µmol	750.0-1500 nmol	5-1030-005	295.00 EUR

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RNA HPLC grade purification and RNA purification

For certain applications RNA HPLC grade purification may be sufficient (but generally PAGE purification is recommended).

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product	scale	cat. no.	price/oligonucleotide
RNA HPLC grade purification	1 µmol	5-1021-004	50.00 EUR
RNA double HPLC	0.2 µmol	5-1024-003	60.00 EUR
	1 µmol	5-1024-004	70.00 EUR
RNA cell culture grade purification	0.2 µmol	5-1023-003	50.00 EUR
	1 µmol	5-1023-004	70.00 EUR
RNA desalting	1 µmol	5-1041-004	20.00 EUR

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DS (desalting) grade purification

This method was developed especially for ultra-short oligonucleotides with a chain length between 1 and 7 bases. It consists of an in-house developed 2 x HPLC purification and a desalting step.

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product	scale	amount	cat. no.	price/oligonucleotide
DS grade purification	0.05 µmol	1 -3 nmol	5-1040-002	30.00 EUR
	0.20 µmol	5 -15 nmol	5-1040-003	38.00 EUR
	1.00 µmol	25-50 nmol	5-1040-004	45.00 EUR
	15.00 µmol	500 -1000 nmol	5-1040-005	550.00 EUR

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Cell culture grade purification

This unique in-house development is a multi-step purification method for all oligonucleotides used in cell culture. It consists of two HPLC purification steps and one sterile filtration step.

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product	scale	amount	cat. no.	price/oligonucleotide
Cell culture grade purification	0.05 µmol	1-3 nmol	5-1050-002	30.00 EUR
	0.20 µmol	5 -15 nmol	5-1050-003	38.00 EUR
	1.00 µmol	25 -50 nmol	5-1050-004	45.00 EUR
	15.0 µmol	500 -1000 nmol	5-1050-005	550.00 EUR

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PAGE grade/FCS grade purification

PAGE is the most powerful method completely removing all types of failure sequences and intercalating dyes like TAMRA. Due to the lower recovery rate IBA NAPS can not guarantee delivery amounts.

Recommended for following products:

• RNA
• Oligonucleotides for biophysical application
• FCS-Spectroscopy

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product	scale	cat. no.	price/oligonucleotide
PAGE purification	0.05 µmol	5-1060-002	100.00 EUR
	0.2 µmol	5-1060-002	125.00 EUR
	1.0 µmol	5-1060-002	150.00 EUR

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Reverse phase C18 HPLC purification

For oligonucleotides requiring more than standard reverse phase purification, such as double dye labeled oligonucleotides.

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product	scale	cat. no.	price/oligonucleotide
Reverse phase C18 HPLC purification	0,05 µmol	5-1022-002	25.00 EUR
	0,2 µmol	5-1022-003	30.00 EUR
	1 µmol	5-1022-004	40.00 EUR

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Documentation

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product	cat. no.	price/oligonucleotide
PAGE documentation	5-1070-000	10.00 EUR
HPLC documentation	5-1070-010	10.00 EUR

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