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OligoÀÇ º¸°ü¹ý ¹× »ç¿ë¹æ¹ý |
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| *Oligonuceotide´Â ÀϹÝÀûÀ¸·Î µ¿°á°ÇÁ¶µÈ pellet»óÅ·Π°ø±ÞµË´Ï´Ù.
À̰ÍÀº ¿î¼Û½Ã, ¿Âµµ µî ¿©·¯°¡ÁöÁ¶°Ç¿¡ ÀÇÇØ ÀϾ¼ö ÀÖ´Â ÇÕ¼ºµÈ DNAµéÀÇ ºÐÇØ¸¦ ¹æÁöÇϱâ À§ÇÔÀÔ´Ï´Ù.
DryµÈ ¿Ã¸®°í´Â ¶§¶§·Î ¿î¼Û°úÁ¤ ¹× ±âŸ»çÀ¯·Î tubeÀÌ ¶Ñ²±À» ¿©´Â¼ø°£ pelletÀÌ ½±°Ô ¼Õ½ÇµÉ¼ö ÀÖ½À´Ï´Ù.
ƯÈ÷ LatexÀå°©µîÀ» ³¢°í ¶Ñ²±À» °³ºÀÇÏ¸é ¸¶Âû¿¡ ÀÇÇØ oligo¸¦ ¼Õ½ÇÇÒ °æ¿ì°¡ ¹ß»ýÇÕ´Ï´Ù.
µû¶ó¼ tubeÀ» ¿±âÀü¿¡ ¸ÕÀú Àá±ñ centrifuge¿¡ µ¹¸®½Ã°í tube¸¦ °³ºÀÇÏ¸é µË´Ï´Ù.
*ÀϹÝÀûÀ¸·Î oligo´Â -20¡É¿¡¼¼ 1³âÁ¤µµ ¾ÈÁ¤ÇÏ´Ù°í º¼¼ö ÀÖ½À´Ï´Ù.
OligoÁ¦Ç°À» ¼ö·ÉÈÄ ¸ê±ÕÁõ·ù¼ö·Î Àß ³ìÀÎÈÄ ¿©·¯ tube¿¡ º¸°üÇϽô°ÍÀÌ ¾ÈÀüÇÕ´Ï´Ù.
³Ãµ¿°ú ÇØµ¿À» ¹Ýº¹Çҽà oligoÀÇ ÀÌ»óÀÌ ¹ß»ýµÉ ¼ÒÁö°¡ ÀÖ½À´Ï´Ù.
¿ë¾×»óÅ·Πº¸°ü½Ã¿¡´Â ¸ê±ÕÁõ·ù¼öº¸´Ù TE(10mM Tri-HCl(Ph8.0), 1mM EDTA)buffer¿¡ ³ìÀ̴°ÍÀÌ ¿À·¡ º¸°üÇÒ ¼ö ÀÖ½À´Ï´Ù.
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| ¢Â µ¿ÀÏÇÑ OligoÇÕ¼º½Ã ¼·Î ´Ù¸¥ OD°ª |
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OLIGOÇÕ¼º¿¡ »ç¿ëµÇ´Â Column³» CPG(controlled pore glass)ÀÇ ¾ç¿¡
µû¶ó ÇÕ¼ºµÇ´Â oligoÀÇ
yiled´Â ´Þ¶óÁý´Ï´Ù.
¿¹¸¦ µé¸é ¼ÒºñÀÚ°¡ 50nmol scale·Î ÁÖ¹®ÇÒ°æ¿ì ÇÕ¼º±âÀÇ column¾È¿¡´Â
ÃÖ¼ÒÇÑ ÇÕ¼ºÀ» À§ÇÑ
3¡¯baseÀÇ ¾çÀÌ 50nmol ÀÌ»óÀÌ ¾çÀ» ³Ö°í ÇÕ¼ºÀ» ½ÃÀÛÇÕ´Ï´Ù.
50nmol scaleÀ̶õ ÇÕ¼º½Ã starting yieldÀ̸ç ÃÖÁ¾ ¾çÀÇ final yield¸¦ ÀǹÌÇϴ°ÍÀº ¾Æ´Õ´Ï´Ù.
* oligo ÇÕ¼º¼öÀ²À» °áÁ¤Áþ´Â ¿äÀÎ:
oligo size, coupling effcency, base
Composition, Á¤Á¦¹æ¹ýµî ÀÔ´Ï´Ù.
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| ¢Â TM(melting temperature)°ª |
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OligonucleotideÀÇ 50%´Â duplexÇüÅÂ, 50%´Â single-strandÇüŸ¦
Áö´Ò¶§ÀÇ ¿Âµµ¸¦ ¸»Çϸç,
oligonudleotideÀÇ sequence, oligonucleotideÀdzóµµ,±×¸®°í
PCR¹ÝÀÀ½ÃÀÇ salt³óµµ µî¿¡ ÀÇÇØ
¿µÇâÀ» ¹Þ½À´Ï´Ù.
* TM°ªÀº Àý´ëÀûÀÎ °ªÀº ¾Æ´Õ´Ï´Ù.
½ÇÇèÀÚ°¡ Template¸¦ ÁغñÇÏ´Â °úÁ¤¿¡¼ »ý±æ¼öÀÖ´Â ¿©·¯°¡Áö ºÒ¼ø¹°°ú saltµé¿¡ ÀÇÇØ ¿©·¯°¡Áö º¯¼ö°¡ »ý±æ¼ö ÀÖ½À´Ï´Ù.
µû¶ó¼ ´ç»ç TM°ªÀ» Âü°í·Î ½ÇÇèÀ» ÇÏ½Ã°í ¸¸¾à ½ÇÇè°á°ú°¡ ³ª¿ÀÁö ¾ÊÀ»°æ¿ì ÇØ´ç TM°ªº¸´Ù 4- 5µµ ³·ÀºÁ¶°ÇÀ¸·Î ½ÇÇèÀ» ÇϽøé ÁÁ½À´Ï´Ù.
* ¸¸ÀÏ ±âÁØTM°ª¿¡¼ Non-specific product°¡ ¸¹Àº °æ¿ì¿¡ annealing
temperature¸¦ ³ô¿©°¡¸é ¼ ½ÇÇèÁ¶°ÇÀ» ã¾Æ¾ß ÇÕ´Ï´Ù.
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| ¢Â ÇÕ¼º Oligo°¡°Ý |
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| ÇÕ¼ºscale |
0.05umol |
0.05umol |
0.05umol |
0.2umol |
1.0umol |
| ÇÕ¼º¿°±â¼ö |
~30mer |
~40mer |
~50mer |
~99mer |
~120mer |
| °¡°Ý/base |
\
650 |
\
650 |
\
750 |
\
1000 |
\
2,500 |
| OPCÁ¤Á¦ |
¹«·á |
¹«·á |
¹«·á |
¹«·á |
¹«·á |
| PAGEÁ¤Á¦ |
\25,000 |
\25,000 |
\25,000 |
\25,000 |
\35,000 |
| HPLCÁ¤Á¦ |
\30,000 |
\30,000 |
\30,000 |
\30,000 |
\50,000 |
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*1ȸ
1,000merÀÌ»ó ÁÖ¹®½Ã 550/mer ÀÔ´Ï´Ù.
*1ȸ 200,000¿ø ÀÌ»ó ÁÖ¹®½Ã ¹®È»óǰ±Ç 1¸Å ¹ß¼Û
*NO Setup charge(´Ü ÁÖ¹®¼ö·®ÀÌ 3column ÀÌÇÏ´Â
Åùè¿ä±Ý
4,000¿ø ºÎ°¡)
**** 50mer ÀÌ»óÀÇ oligoÇÕ¼ºÀÇ °æ¿ì PAGE/HPLCÁ¤Á¦¸¦
±ÇÇØµå¸³´Ï´Ù. ****
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| ¢Â Modification Oligo List |
| 5¡¯
Fluorescein |
3¡¯
Fluorescein |
5¡¯
Biotin |
3¡¯
Biotin |
| 5¡¯
Phosphate |
3¡¯
Phosphate |
5¡¯
Amine(C6) |
3¡¯
Amine(C6) |
| 5¡¯
TAMRA |
3¡¯
TAMRA |
5¡¯
Thiol(C6) |
3¡¯
Thiol(C3) |
| 5¡¯
Rhodamine |
3¡¯
Rhodamine |
5¡¯
Dig |
3¡¯
Dig |
| 5¡¯
FAM |
5¡¯
HEX |
5¡¯
TET |
5¡¯
Texas Red |
2¡¯-
O - Methyl
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Taqman
Probe |
Phosphorothioate |
RNA |
| Inosine(dl) |
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| *
´ë·®ÇÕ¼º ¹× ±âŸ Modification oligo ´Â ¹®Àǹٶø´Ï´Ù.
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