To avoid polymerase extension at the 3’ terminus, 2’,3’-dideoxynucleoside and 3’-deoxynucleoside CPGs have proved to be effective. 2’,3’-phosphoramidites are designed to be used with the 5’-phosphoramidites and supports. Since these phosphoramidites have no DMT group, they are not compatible with purification by the DMT-on technique. Ion exchange HPLC or PAGE should be used to purify these dideoxy terminated oligos to ensure that shorter sequences (containing 3’-OH) groups are removed. (3’-termination can also be effected using a 3’-3’ linkage formed using 5’-supports, or 3’-spacer C3 CPG.)
DILUTION/COUPLING DATA
The table below shows pack size data and, for solutions, dilutions and approximate couplings based on normal priming procedures.
ABI 392/394 |
Cat.No. |
Pack Size |
Grams/ Pack |
0.1M Dil. (mL) |
LV40 |
LV200 |
40nm |
0.2µm |
1µm |
10µm |
Approximate Number of Additions |
10-7301-02 |
0.25grams |
.25grams |
5.86 |
182 |
109.2 |
68.25 |
49.64 |
36.4 |
9.1 |
10-7301-90 |
100µmoles |
.043grams |
1 |
20 |
12 |
7.5 |
5.45 |
4 |
1 |
Expedite |
Cat.No. |
Pack Size |
Grams/ Pack |
Dilution (mL) |
Molarity |
50nm |
0.2µm |
1µm |
15µm |
|
Approximate Number of Additions |
10-7301-02 |
0.25grams |
.25grams |
8.75 |
.07 |
168.6 |
105.38 |
76.64 |
10.54 |
|
10-7301-90 |
100µmoles |
.043grams |
1.5 |
.07 |
23.6 |
14.75 |
10.73 |
1.48 |
|
Beckman |
Cat.No. |
Pack Size |
Grams/ Pack |
Dilution (mL) |
Molarity |
30nm |
200nm |
1000nm |
|
|
Approximate Number of Additions |
10-7301-02 |
0.25grams |
.25grams |
8.75 |
.07 |
170.2 |
106.38 |
77.36 |
|
|
10-7301-90 |
100µmoles |
.043grams |
1.5 |
.07 |
25.2 |
15.75 |
11.45 |
|
|
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