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³óµµ
- Plasmid : 100ng/ul, PCR
product(purified) : 50ng/ul, 1.5mL Microcentrifuge
tube¸¦ ±ÇÀåÇÕ´Ï´Ù.
volume
- Plasmid, PCR product °øÅë ±âº» 5ul, ÇÑ ¹æÇâ ½ÇÇè´ç 2ul Ãß°¡
ex) sample 1°³¸¦ »ç¿ëÇÏ¿© 4reaction ½Ã 5ul + (2ul * 4) = 13ul
* ¿î¼ÛµµÁß sampleÀÇ dry·Î ÀÎÇÏ¿© ¾à 10~20% ¼Õ½ÇÀ» °¨¾ÈÇØ
* ÃæºÐÇÑ volumeÀ» º¸³»ÁÖ¼¼¿ä
* Àç¹ÝÀÀ°ú Ãß°¡¹ÝÀÀÀº 1reaction ´ç 5ul, 3ul Ãß°¡
16S-rSequeningÀ» ÀÇ·Ú½Ã,
(cell) agar plate¿¡ colony°¡ º¸ÀÏÁ¤µµ·Î Ű¿îÈÄ¿¡ º¸³»Áֽøé
Single colony¸¦ picking ÇÏ¿© ±ú²ýÇÑ °á°ú°¡
³ª¿Ã¼ö ÀÖ½À´Ï´Ù.
glycerolº¸´Ù agar·Î º¸³»Áֽô°ÍÀÌ ºü¸£°í ±ú²ýÇÑ °á°ú°¡ ³ª¿É´Ï´Ù.
(gDNA)·Î º¸³»ÁÖ´Â °æ¿ì, ³óµµ 30-50ng/uL, 5-10uL ÀÌ»óÀ¸·Î º¸³»ÁÖ½Ã¸é µË´Ï´Ù.
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5
sample ±âÁØ, 5 pmole/ul (5uM)
³óµµ·Î 20ul
Reaction
¼ö°¡ Ãß°¡µÉ ¶§¸¶´Ù 10 pmole ¾¿ Áõ°¡½ÃÄÑ
ÃæºÐÇÑ ¾çÀ» µ¿ºÀÇØÁֽʽÿÀ.
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primer Á¦°ø ½Ã »ùÇðú ºÐ¸®µÈ ¿ë±â
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¼ºñ½º¿¡ ÀûÇÕÇÑ Primer Tm : 50~58
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