Glen UnySUPPORT

Our original Universal Support (20-5000) has been discontinued since complete dephosphorylation using ammonium hydroxide, AMA or anhydrous methylamine gas takes longer than most companies wish to allocate. A recent development has been the use of a support based on a molecule which is "conformationally preorganized" to accelerate the dephosphorylation reaction.1,2 By using a rigid bicyclic molecule on the support, the rate of elimination is markedly faster than the original Universal Support. The structure of Glen UnySupport is shown below. The N-phenyl version, developed at Isis Pharmaceuticals as UnyLinker™ , is available from several companies for large scale oligo synthesis. Glen UnySupport is the N-methyl version, which is preferred for high throughput oligonucleotide synthesis since methylamine rather than aniline is formed on deprotection. We are happy to offer Glen UnySupport in a variety of popular formats under license from Isis Pharmaceuticals.
Item Catalog No. Pack Price ($)
Bulk Supports
20-5040-01 0.1g 11.00
  (500Å CPG) 20-5040-02 0.25g 25.00
20-5040-10 1.0g 95.00
20-5041-01 0.1g 11.00
20-5041-02 0.25g 25.00
20-5041-10 1.0g 95.00
25-5040-01 0.1g 15.00
25-5040-02 0.25g 30.00
25-5040-10 1.0g 115.00
26-5040-01 0.1g 16.00
26-5040-02 0.25g 35.00
26-5040-10 1.0g 125.00
Columns
  The 1000Å columns and frits below are routinely stocked.
ABI Format (not LV)
20-5141-41 Pack of 4 60.00
  0.2 µmole columns 20-5141-42 Pack of 4 40.00
  40 nmole columns 20-5141-45 Pack of 4 40.00
  10 μmole column (TWIST Format) 20-5141-13 Pack of 1 100.00
  40 nmole frits 20-5441-95 96x40nm 150.00
20-0060-00 Pack of 10 20.00
AB 3900 Format
Glen UnySupport PS
26-5140-52 Pack of 10 100.00
  40 nmole columns 26-5140-55 Pack of 10 100.00
Expedite Format
20-5241-41 Pack of 4 60.00
  0.2 µmole columns 20-5241-42 Pack of 4 40.00
  40 nmole columns 20-5241-45 Pack of 4 40.00
  15 μmole column (TWIST Format) 20-5241-14 Pack of 1 150.00
96 Well Format (MerMade, etc.)
20-5141-91 Pack of 96 375.00
  200 nmole columns 20-5141-92 Pack of 96 250.00
  40 nmole columns 20-5141-95 Pack of 96 250.00
Glen UnySupport™ 500
Glen UnySupport™ 500
Glen UnySupport™ PS
Glen UnySupport™ PS
Glen UnySupport™ 1000
Glen UnySupport™ 1000
High Load Glen UnySupport™
High Load Glen UnySupport™

ELIMINATION CONDITIONS

Reagent
Conditions
Ammonium
hydroxide
80°C/2h
55°C/8h
Ammonium
hydroxide/40%
Methylamine(AMA)
80°C/0.5h
65°C/1h
55°C/8h
Methylamine Gas 65°C/0.5h/30psi
Potassium
Carbonate
in Methanol
RT/17h
t-Butylamine/
Water (1:3 v/v)
60°C/4h

Intellectual Property

These products are covered by US Patent 7,202,264 owned by Ionis Pharmaceuticals, Inc..

Reference(s)

  1. A.P. Guzaev, and M. Manoharan, J Am Chem Soc, 2003, 125, 2380- 2381.
  2. R.K. Kumar, A.P. Guzaev, C. Rentel, and V.T. Ravikumar, Tetrahedron, 2006, 62, 4528.

OTHER INSTRUMENT TYPES

All minor bases, RNA products and modifiers are packaged in septumcapped vials suitable for ABI and other instruments. If you would like another type of vial/column add the following to the end of the catalog number.

Monomers  

For Instrument type
Add
Expedite
E
Mermade
M

Columns  

For Instrument type
Add
Expedite
E
Applied Biosystems 3900
A
Mermade
M

Please inquire for availability of columns for other instrument types.

Glen UnySupport FC

The extended time required to cleave the succinate linkage of the original Glen UnySupport can be problematical, especially in high-throughput production of oligos, due to the outgassing of ammonia and/or methylamine. This reduction in concentration of gas can necessitate the evaporation of the cleavage solution and addition of fresh Ammonium Hydroxide:MethylAmine 1:1 (AMA) or ammonium hydroxide (NH4OH) to ensure complete deprotection and dephosphorylation of the product oligos. Using a diglycolate linkage in Glen UnySupport FC instead of the succinate in Glen UnySupport, a significant increase in the rate of cleavage has been achieved. The minimum cleavage times for both versions are as follows:


AMANH4OH
Glen UnySupport10 min.40 min.
Glen UnySupport FC2 min.5 min.

With the cleavage time of Glen UnySupport FC reduced to less than 5 minutes, there is minimal loss of volatile gas and, therefore, no need to evaporate the cleavage solution and replenish with fresh AMA or ammonium hydroxide solutions.

We offer Glen UnySupport FC attached to 1000Å CPG in a variety of formats suited to high throughput synthesis, as well as in bulk for more routine use.

Item Catalog No. Pack Price ($)
Bulk Support
22-5041-01 0.1g 11.00
  (1000Å CPG) 22-5041-02 0.25g 25.00
22-5041-10 1.0g 95.00
ABI Format (not LV)
22-5141-41 Pack of 4 60.00
  0.2 µmole columns 22-5141-42 Pack of 4 40.00
  40 nmole columns 22-5141-45 Pack of 4 40.00
  10 μmole column (TWIST Format) 22-5141-13 Pack of 1 100.00
AB 3900 Format
22-5141-52 Pack of 10 100.00
  40 nmole columns 22-5141-55 Pack of 10 100.00
Expedite Format
22-5241-41 Pack of 4 60.00
  0.2 µmole columns 22-5241-42 Pack of 4 40.00
  40 nmole columns 22-5241-45 Pack of 4 40.00
  15 μmole column (TWIST Format) 22-5241-14 Pack of 1 150.00
96 Well Format (MerMade, etc.)
22-5141-91 Pack of 96 375.00
  0.2 µmole columns 22-5141-92 Pack of 96 250.00
  40 nmole columns 22-5141-95 Pack of 96 250.00
Glen UnySupport™ FC
Glen UnySupport™ FC

ELIMINATION CONDITIONS

Reagent
Conditions
Ammonium hydroxide 80°C/2h
55°C/8h
Ammonium hydroxide/40% Methylamine (AMA) 80°C/0.5h
65°C/1h
55°C/8h
Methylamine Gas 65°C/0.5h/30psi
Potassium Carbonate RT/17h in Methanol
t-Butylamine/Water (1:3 v/v) 60°C/4h

Intellectual Property

22-5041 is covered by US Patent 7,202,264 owned by Ionis Pharmaceuticals, Inc..

Universal Support III

The key step in the use of any universal support in oligonucleotide synthesis is the dephosphorylation of the 3'-phosphate group to form the desired 3'-hydroxyl group. Azhayev1,2 has excelled in the investigation of neighboring group assistance in the dephosphorylation reaction. Amide groups may be considered to be weak N‑H acids and can display basic properties in ammonium hydroxide or aqueous methylamine. In the original work1,2 (±)-3-Amino-1,2-propanediol was used to form a novel universal support(1). A succinate linker attaches the 3-amino group to the support and the 2-OH is protected with a base-labile group to set up an amide assisted elimination in mildly basic conditions. In this way, the dephosphorylation reaction would eliminate the desired 3'-OH oligonucleotide into solution and the product of any β-elimination competing side reaction would remain bound to the support. A further improvement has been achieved by using a carbamate group to connect the universal linker to the support, now called Universal Support III. The structures of the two supports are shown below right. Because the universal linker is unchanged and the succinate or carbamate groups remain attached to the support, as in product Universal Support III (2). Using Universal Support III, an oligo yield of > 80% can be achieved on polymeric supports, with purity equivalent to the same oligo prepared normally.

Conditions for Cleavage and Deprotection are outlined in the table opposite. Universal Support III has been shown to generate oligonucleotides with the same efficacy in polymerase extension reactions as regular oligos. Despite the mild elimination reaction, oligonucleotides up to 75mer in length can be prepared routinely without loss of oligo during the synthesis cycles. This support is also used for the production of siRNA oligos.
Item Catalog No. Pack Price ($)
26-5010-01 0.1g 16.00
26-5010-02 0.25g 35.00
26-5010-10 1.0g 125.00
ABI Format (not LV)
Universal Support III PS
26-5110-41 Pack of 4 60.00
  0.2 µmole columns 26-5110-42 Pack of 4 40.00
  40 nmole columns 26-5110-45 Pack of 4 40.00
  10 μmole column (TWIST Format) 26-5110-13 Pack of 1 100.00
Expedite Format
26-5210-41 Pack of 4 60.00
  0.2 µmole columns 26-5210-42 Pack of 4 40.00
  40 nmole columns 26-5210-45 Pack of 4 40.00
  15 μmole column (TWIST Format) 26-5210-14 Pack of 1 150.00
96 Well Format (MerMade, etc.)
Universal Support III PS
26-5110-91 Pack of 96 375.00
  200 nmole columns 26-5110-92 Pack of 96 250.00
  40 nmole columns 26-5110-95 Pack of 96 250.00
AB 3900 Format
Universal Support III PS
26-5110-52 Pack of 10 100.00
  40 nmole columns (AB 3900 Format) 26-5110-55 Pack of 10 100.00
Universal Support III PS
Universal Support III PS

CLEAVAGE AND DEPROTECTION

  1. Cleavage
    For standard and UltraFast deprotection protocols, cleave the oligo from the support using 2M ammonia in methanol at room temperature for 30 minutes. (Only for oligonucleotides greater than 50 nucleotides in length, rinse the support with a further volume of water. Combine the two washes and evaporate to dryness.)
  2. Deprotection
    Standard
    Add 1 volume of 30% ammonium hydroxide, seal and deprotect using the conditions appropriate for removal of the protecting groups on the nucleobases.

    UltraFast
    Add 1 volume of AMA (ammonium hydroxide/40% aqueous methylamine 1:1) seal and deprotect at 65°C for 10 minutes.

    UltraMild
    Using Ammonium Hydroxide
    Add 1 volume of ammonium hydroxide, seal and leave at room temperature for 8 hours.
  3. UltraMild Cleavage and Deprotection
    Using Potassium Carbonate in Methanol
    Cleave the oligo from the support using 50 mM potassium carbonate in methanol at room temperature for 30 minutes. Seal and leave overnight at room temperature.

Intellectual Property

26-5010 is covered by US Patent Nos.: 6,770,754 and 7,491,817 and European Patent Nos.: 1404695 and 2248820.

Reference(s)

  1. A.V. Azhayev, Tetrahedron, 1999, 55, 787-800.
  2. A.V. Azhayev and M. Antopolsky, Tetrahedron, 2001, 57, 4977-4986.

UNIVERSAL HybridCPG SOLID SUPPORTS

HybridCPG™ consists of CPG particles conformally coated with a very thin crosslinked polymer film based on polystyrene. The molecular structure of the polymer coating is designed to have a very high density of evenly distributed attachment points for optimum oligo synthesis. In this way, the pore size to loading density trade-off is minimized and the chemical resistance to glass-aggressive reagents is greatly improved. Although the nano-scale coating is subject to swelling, the rigid pore structure of the CPG substrate accommodates this and, at the same time, maintains a uniform pore space for the oligo synthesis. Thus, HybridCPG exhibits no bulk swelling in the synthesis solvents and allows much higher ligand loadings for a given pore size compared to conventional CPG. Glen Research is partnering with Prime Synthesis, developers of HybridCPG, to introduce our two popular Universal Supports, Glen UnySupport™ and Universal Support III, on HybridCPG.
Item Catalog No. Pack Price ($)
Bulk Support
28-5010-01 0.1g Discontinued
28-5010-02 0.25g Discontinued
28-5010-10 1.0g Discontinued
  Universal Support III HybridCPGTM has been discontinued.
28-5040-01 0.1g 16.00
28-5040-02 0.25g 35.00
28-5040-10 1.0g 125.00
Glen UnySupportTM HybridCPGTM
Glen UnySupportTM HybridCPGTM

TRADEMARKS

HybridCPG™ is a trademark of Prime Synthesis, Inc. Glen UnySupport™ is a trademark of Glen Research Corporation.

Intellectual Property

Glen UnySupportTM HybridCPGTM is covered by US Patent 7,202,264 owned by Ionis Pharmaceuticals, Inc..

Universal Support III HybridCPGTM is covered by US Patent Nos.: 6,770,754 and 7,491,817 and European Patent Nos.: 1404695 and 2248820.

Q-SUPPORTS

Oligonucleotides are routinely prepared on supports to which the first nucleoside is attached via a succinate linkage. Over the years, the succinate linkage has demonstrated stability during the synthesis process but has sufficient lability to be cleaved quickly in the deprotection step. However, if the cleavage step is carried out with ammonium hydroxide manually or on the synthesizer, it consumes one hour of precious time while releasing only about 80% of the oligonucleotide. This step is, therefore, a bottleneck in the productivity of many synthesis groups.

Is it possible to find a replacement to the succinate group which offers good stability to the synthesis reagents while offering a much faster cleavage step? The oxalate group has been shown to be very labile during cleavage but its stability to the normal synthesis reagents is not good, requiring changes for successful use. In a practical but elegant study1 of various bifunctional carboxylic acids, Richard Pon's group identified hydroquinone-O,O'-diacetic acid as the most satisfactory alternative to the succinate group. Nucleosides with this linker arm (Q-linker) are attached to supports with the same ease as the succinyl linker arm.

The cleavage time in ammonium hydroxide at room temperature was found to be 2 minutes, but what about the stability during synthesis? How significant was premature cleavage of oligonucleotide on the synthesizer because of the basic reagents in the capping mixes and oxidizer? Pon showed that the Q-linker is stable to the capping reagents but very slightly labile to the oxidizer (8% cleavage in overnight exposure which would correspond to about 2,000 normal synthesis cycles).

We tested the significance of premature cleavage by preparing sixteen 20mer oligonucleotides on a 0.2 µmole scale, eight with succinate and eight with Q-linkers. The succinate supported oligos were cleaved for 1 hour at room temperature, while those on the Q-support were cleaved for 2 minutes. Both sets were then deprotected normally with ammonium hydroxide. The Q-supports actually gave 5% better yields of product than the succinate supports. Oligo purities were equivalent in both sets.

The Q-linker is absolutely compatible with all hydrolytic cleavage procedures, but especially mild procedures like potassium carbonate in methanol. Pon also showed that it is preferable for RNA supports, improving the cleavage time for 2'-silyl protected nucleoside supports from 2 hours (60-65% cleavage) to 5 minutes (95% cleavage).

We are offering Q-linkers of the four regular nucleosides on 500Å CPG in 0.2 and 1µmole scales.

Oligonucleotides are routinely prepared on supports to which the first nucleoside is attached via a succinate linkage. Over the years, the succinate linkage has demonstrated stability during the synthesis process but has sufficient lability to be cleaved quickly in the deprotection step. However, if the cleavage step is carried out with ammonium hydroxide manually or on the synthesizer, it consumes one hour of precious time while releasing only about 80% of the oligonucleotide. This step is, therefore, a bottleneck in the productivity of many synthesis groups.

Is it possible to find a replacement to the succinate group which offers good stability to the synthesis reagents while offering a much faster cleavage step? The oxalate group has been shown to be very labile during cleavage but its stability to the normal synthesis reagents is not good, requiring changes for successful use. In a practical but elegant study1 of various bifunctional carboxylic acids, Richard Pon's group identified hydroquinone-O,O'-diacetic acid as the most satisfactory alternative to the succinate group. Nucleosides with this linker arm (Q-linker) are attached to supports with the same ease as the succinyl linker arm.

The cleavage time in ammonium hydroxide at room temperature was found to be 2 minutes, but what about the stability during synthesis? How significant was premature cleavage of oligonucleotide on the synthesizer because of the basic reagents in the capping mixes and oxidizer? Pon showed that the Q-linker is stable to the capping reagents but very slightly labile to the oxidizer (8% cleavage in overnight exposure which would correspond to about 2,000 normal synthesis cycles).

We tested the significance of premature cleavage by preparing sixteen 20mer oligonucleotides on a 0.2 µmole scale, eight with succinate and eight with Q-linkers. The succinate supported oligos were cleaved for 1 hour at room temperature, while those on the Q-support were cleaved for 2 minutes. Both sets were then deprotected normally with ammonium hydroxide. The Q-supports actually gave 5% better yields of product than the succinate supports. Oligo purities were equivalent in both sets.

The Q-linker is absolutely compatible with all hydrolytic cleavage procedures, but especially mild procedures like potassium carbonate in methanol. Pon also showed that it is preferable for RNA supports, improving the cleavage time for 2'-silyl protected nucleoside supports from 2 hours (60-65% cleavage) to 5 minutes (95% cleavage).

We are offering Q-linkers of the four regular nucleosides on 500Å CPG in 0.2 and 1µmole scales.

Catalog No. Catalog No. Catalog No. Catalog No. Catalog No. Pack Price($)
dA dC dT Ac-dC dmf-dG    
500Å Bulk Support
21-2000-01 21-2010-01 21-2030-01 21-2013-01 21-2029-01 0.1g 11.00
21-2000-02 21-2010-02 21-2030-02 21-2013-02 21-2029-02 0.25g 25.00
21-2000-10 21-2010-10 21-2030-10 21-2013-10 21-2029-10 1.0g 95.00
ABI Format (not LV)
21-2100-41 21-2110-41 21-2130-41 21-2113-41 21-2129-41 Pack of 4 60.00
21-2100-42 21-2110-42 21-2130-42 21-2113-42 21-2129-42 Pack of 4 40.00
Expedite Format
21-2200-41 21-2210-41 21-2230-41 21-2213-41 21-2229-41 Pack of 4 60.00
21-2200-42 21-2210-42 21-2230-42 21-2213-42 21-2229-42 Pack of 4 40.00
dA-Q-CPG 500
dA-Q-CPG 500
dC-Q-CPG 500
dC-Q-CPG 500
dT-Q-CPG 500
dT-Q-CPG 500
Ac-dC-Q-CPG 500
Ac-dC-Q-CPG 500
dmf-dG-Q-CPG  500
dmf-dG-Q-CPG 500

Q/SUCCINATE COMPARISON

Q-Support
Succinate
(2 minutes
(60 minutes
cleavage)
cleavage)
132 ODU*
125 ODU*

*Average crude yield from eight 1µmole columns deprotected normally.

Reference(s)

  1. (1) R.T. Pon and S.Y. Yu, Tetrahedron Lett, 1997, 38, 3327-3330.

OTHER INSTRUMENT TYPES

All minor bases, RNA products and modifiers are packaged in septumcapped vials suitable for ABI and other instruments. If you would like another type of vial/column add the following to the end of the catalog number.

Monomers  

For Instrument type
Add
Expedite
E
Mermade
M

Columns  

For Instrument type
Add
Expedite
E
Applied Biosystems 3900
A
Mermade
M

Please inquire for availability of columns for other instrument types.

HIGH LOAD CPG

Our high loading support is based on controlled pore silica and it retains the usual 500Å pores. The spacer is also conventional. The only significant difference is the loading which is in the range 80 - 130µmoles/g or about 2.5 times the loading of normal 500Å CPG. Typical loadings for our high load CPG are in the 100 - 120µmoles/g range. As a consequence of the high loading, this support should not be used for sequences longer than 40mers. This high loading support is available in columns for most synthesizers. The 2.5µmole column is identical to our standard 1µmole column (with the exception of the loading). It should be used on occasions when greater than 1µmole is desired but when a 10 or 15µmole synthesis is too high. It should be run using the 1µmole cycle. The 25µmole column is identical to the 10µmole column used on Applied Biosystems synthesizers. It is run using the 10µmole cycle. The 35µmole column is used as an alternative to the 15µmole Expedite column. Again no changes to the standard cycle are recommended. The support is of course available in bulk for use on large-scale synthesizers. A word of caution is in order. When using a column with a higher load than recommended by the instrument manufacturer, there is a much smaller margin for error. All reagents must be fresh and anhydrous diluent and activator must be used. Should you decide to prepare higher-loading columns, ensure that the molar excess of monomer to support nucleoside is at least 5X and preferably 10X.

Our high loading support is based on controlled pore silica and it retains the usual 500Å pores. The spacer is also conventional. The only significant difference is the loading which is in the range 80 - 130µmoles/g or about 2.5 times the loading of normal 500Å CPG. Typical loadings for our high load CPG are in the 100 - 120µmoles/g range. As a consequence of the high loading, this support should not be used for sequences longer than 40mers. This high loading support is available in columns for most synthesizers. The 2.5µmole column is identical to our standard 1µmole column (with the exception of the loading). It should be used on occasions when greater than 1µmole is desired but when a 10 or 15µmole synthesis is too high. It should be run using the 1µmole cycle. The 25µmole column is identical to the 10µmole column used on Applied Biosystems synthesizers. It is run using the 10µmole cycle. The 35µmole column is used as an alternative to the 15µmole Expedite column. Again no changes to the standard cycle are recommended. The support is of course available in bulk for use on large-scale synthesizers. A word of caution is in order. When using a column with a higher load than recommended by the instrument manufacturer, there is a much smaller margin for error. All reagents must be fresh and anhydrous diluent and activator must be used. Should you decide to prepare higher-loading columns, ensure that the molar excess of monomer to support nucleoside is at least 5X and preferably 10X.

Item Catalog No. Catalog No. Catalog No. Catalog No. Pack Price($)
dA dC dG dT    
Columns
25-2100-46 25-2110-46 25-2120-46 25-2130-46 4x2.5µm 75.00
25-2100-17 25-2110-17 25-2120-17 25-2130-17 1x25µm 125.00
(Expedite) 25-2200-46 25-2210-46 25-2220-46 25-2230-46 4x2.5µm 75.00
25-2200-18 25-2210-18 25-2220-18 25-2230-18 1x35µm 185.00
Bulk
25-2000-02 25-2010-02 25-2020-02 25-2030-02 0.25g 25.00
25-2000-10 25-2010-10 25-2020-10 25-2030-10 1.0g 90.00
dA-High Load-CPG
dA-High Load-CPG
dC-High Load-CPG
dC-High Load-CPG
dG-High Load CPG
dG-High Load CPG
dT-High Load-CPG
dT-High Load-CPG

OTHER INSTRUMENT TYPES

All minor bases, RNA products and modifiers are packaged in septumcapped vials suitable for ABI and other instruments. If you would like another type of vial/column add the following to the end of the catalog number.

Monomers  

For Instrument type
Add
Expedite
E
Mermade
M

Columns  

For Instrument type
Add
Expedite
E
Applied Biosystems 3900
A
Mermade
M

Please inquire for availability of columns for other instrument types.

See Also


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