5' MMT-Amino C6 Modifier (MMT-6-Aminohexyl Amidite)
货号：BNS-5015-100,BNS-5015-250,BNS-5015-50
规格：100 µmol,250 mg,50 µmol
价格：844,3236,450
产品类型：核酸合成
品牌：LGC Biosearch Technologies
| This compound is typically used to add amino functionality to oligonucleotides. For best results leave the MMT on the oligonucleotide during ammonia deprotection. Subsequent cleavage of the MMT group can be accomplished by solution phase methods using 2.5% aqueous TFA or by column cartridge purification (see product usage). |
| 特性： |
| - Chemical Name: 6-(4-Monomethoxytritylamino)hexyl-(2-cyanoethyl)-
(N,N-diisopropyl)-phosphoramidite - Formula: C35H48N3O3P - Molecular Weight: 589.75 - Appearance: colorless oil |
| 用途： |
Synthesis conditions:Prior to dilution ensure that all product is at the bottom of the vial. Dilute to the recommended concentration and mix thoroughly in the sealed vial to ensure that all contents are dissolved.Dilution:100 µmol/mLCoupling:2-minute couplingCleavage conditions:This product is compatible with concentrated ammonium hydroxide or t-butylamine for cleavage. The MMT group may be left on or removed prior to cleavage of the oligo. We recommend leaving the MMT group on the oligo to prevent a side reaction resulting in the elimination of the amine linkage during cleavage.Deprotection conditions: If using fast deprotecting amidites, deprotect in concentrated NH4OH for 1 hour at 60°C. If using standard amidites; cleave in concentrated NH4OH for 5 hours at 60°C. If all the solution is not used during the synthesis, it can be stored under argon and its functionality maintained up to 48 hours.
Kinetics for the removal of the MMT group are slow. To ensure complete removal of the MMT group we suggest the following conditions:
For removal of the MMT group on the DNA synthesizer: Use 3% TCA (Trichloroacetic acid) in DCM and leave on for 5 minutes or use 3% DCA/DCM and leave on for 10 minutes. Cleave as normal.
For removal of MMT group using reversed-phase cartridge purification: Oligo should be cleaved and MMT group left on for this purification procedure. Use a reversed-phase MicroPure column (MP-1602) for cleanup of 5' MMT oligonucleotides. The TFA step cleaves the MMT group resulting in a completely deprotected oligonucleotide. Below is a protocol using reverse phase MicroPure column (MP-1602) for cleanup of 5' MMT oligonucleotides. Each MicroPure column will clean up to a 1µmol synthesis column. Note that the TFA step cleaves the MMT group resulting in a completely deprotected oligonucleotide.
1. Rinse column with CH3CN (2 x 2 mL).
2. Activate column with 1N TEAA (3 x 2 mL)
3. Add Crude MMT-Oligo 2 mL of 1:1 solution (NH4OH/H2O). (Collect and reload solution twice)
4. Elute Failures with 3% NH4OH (3 x 2 mL).
5. Rinse column with water (3 x 2 mL).
6. Deblock MMT with 2.5% TFA (3 x 2 mL). Wait 10 minutes between applications.
7. Rinse excess TFA with water H2O. (3 x 2 mL; rinse until pH is neutral).
8. Insert collection tubes; elute purified oligo with 20% MeCN in water. (1 x 1.5 mL)
9. Dry sample in speed vac
Preparation of Reagents
Five different solutions for the purification process need to be prepared. To store these solutions use clean glass bottles with plastic lined caps.
1. Water. Use good quality sterile, deionized water. HPLC grade is best, especially if subsequent analyses will rely on HPLC methods.
2. Acetonitrile (VWR, cat. number JT9018-3).
3. 20% acetonitrile in water. Dilute 20 mL of the same acetonitrile as in 2 above with 80 mL of the same water as in 1 above.
4. 1.0 N Triethylammonium acetate (TEAA). For this solution, place about 60 mL of the same water as in 1 above in a clean bottle, and add exactly 5.7 mL of acetic acid (Aldrich cat. number 24, 285-3; Caution - causes burns, wear eye protection and gloves when using concentrated acetic acid). Swirl the solution until completely mixed, and add exactly 13.9 mL of triethylamine (Aldrich cat. number 23, 962-3; Caution - wear eye protection and gloves when using concentrated triethylamine). Mix the solution until it is completely homogeneous - a small stir bar and magnetic stirrer may be used with good results. After complete mixing is achieved, the pH should be neutral. pH paper can be used for this (VWR cat. number 60775-702).
5. 2.5% Trifluoroacetic acid (TFA) in water. Carefully add 2.5 mL of TFA (Aldrich cat. number T6, 220-0 Caution - wear eye protection and gloves when using concentrated TFA) to 97.5 mL of the same water as in 1 above. Mix the solution until it is completely homogeneous.
6. 3.0% Ammonium Hydroxide in water. Add 2.8 mL of concentrated ammonia (Aldrich cat. number 22, 122-8 Caution - wear eye protection and gloves when using concentrated ammonia) to 97 mL of the same water as in 1 above. Mix the solution until it is completely homogeneous.Image of cleaved and deprotected structure: The mass this product adds after conjugation and work-up (the additional mass seen by mass spectrometry) is:179.15 |
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