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cDNA Library Construction Kit

-

Á¦Á¶»ç Á¦Ç°ÄÚµå Á¦Ç°¸í ¿ë·® °¡°Ý
(ºÎ°¡¼¼º°µµ)
ºñ°í »ç¿ëÀڸŴº¾ó
Clontech
634901
SMART cDNA Library Construction Kit
°ü·ÃÇмú ±¸¸ÅÇϱâ LMOÁ¦Ç° 
1 Set
2,226,000¿ø  Á¦Á¶»ç ÆäÀÌÁö·Î ¹Ù·Î°¡±â x33044, x33045, x33612
Clontech
635002
pRetro-Lib Vector
°ü·ÃÇмú ±¸¸ÅÇϱ⠶óÀ̼±½º 
20 ¥ìg
657,000¿ø  Á¦Á¶»ç ÆäÀÌÁö·Î ¹Ù·Î°¡±â x33005, x33530
Clontech
635003
pEXP-Lib Vector
°ü·ÃÇмú ±¸¸ÅÇϱâ
20 ¥ìg
705,000¿ø  Á¦Á¶»ç ÆäÀÌÁö·Î ¹Ù·Î°¡±â x33363

  • ÀûÀº ¾çÀÇ total RNA ¶Ç´Â poly A+ RNA·ÎºÎÅÍ °íǰÁúÀÇ cDNA library Á¦ÀÛ
  • ÃÖ¼Ò 50 ngÀÇ total RNA¿¡¼­ cDNA ÇÕ¼º °¡´É
  • µ¶Ã¢ÀûÀÎ SMART cDNA ÇÕ¼º¹ýÀ¸·Î full-length cDNA¸¦ Æ÷ÇÔÇÏ´Â library Á¦ÀÛ
  • Adaptor ligation °úÁ¤ ºÒÇÊ¿ä

SMART cDNA Library Construction Kit´Â phage ¥ëTriplEx2 vector¿¡ full-length cDNA¸¦ cloningÇÏ´Â Á¦Ç°ÀÌ´Ù. ÀÌ Á¦Ç°Àº cDNAÁõÆøÀ» À§ÇÑ SMART ±â¼ú°ú adaptor ¾øÀÌ ¥ëTriplEx2 vector¿¡ cloningÇÏ´Â ±â¼úÀ» ÀÌ¿ëÇϸç ÃÊ±â ½Ã·á ¾ç¿¡ µû¶ó µÎ °¡Áö ½ÇÇè °úÁ¤À¸·Î ³ª´²Á® ÀÖ´Ù. ù ¹øÂ° °úÁ¤Àº »õ·Ó°Ô °í¾ÈµÈ long-distance PCR(LD-PCR) ¹æ¹ýÀ¸·Î 50 ngÁ¤µµÀÇ ÀûÀº ¾çÀÇ RNA ½Ã·á¿¡ Àû¿ëÇÒ ¼ö ÀÖ´Ù (1). µÎ ¹øÂ° °úÁ¤Àº Á» ´õ ÀϹÝÀûÀÎ °úÁ¤À¸·Î ºñ±³Àû ¸¹Àº ¾çÀÇ Ãʱ⠽÷á (¿¹¸¦ µé¸é, 1 ¥ìg ¶Ç´Â ±× ÀÌ»óÀÇ poly A+ RNA)À» »ç¿ëÇÏ´Â ½ÇÇèÀÚ¿¡°Ô ÀûÇÕÇÏ´Ù. µÎ °úÁ¤ ¸ðµÎ ¸¹Àº ¾çÀÇ full-lengthÀÇ double-strand cDNA¸¦ ÇÕ¼ºÇϱâ À§ÇÏ¿© 1st strand ÇÕ¼º°úÁ¤¿¡¼­ SMART IV Oligonucleotide¸¦ ÀÌ¿ëÇÑ´Ù. µÎ ½ÇÇè °úÁ¤ ¸ðµÎ UserManual¿¡ Ç¥±âµÇ¾î ÀÖÀ¸¸ç °¢ÀÚÀÇ ½ÇÇè¿¡ ¸Â°Ô ¼±ÅÃÇϵµ·Ï ÇÑ´Ù. °¢°¢ÀÇ kit´Â cDNA library 7ȸ Á¦ÀÛ ºÐÀÌ´Ù.
¶ÇÇÑ, Æ÷À¯·ù ¹ßÇö¿ë vectorÀÎ pEXP-Lib, retroviral expression vectorÀÎ pRetro-Lib µî SMART cDNA Library Construction Kit¿Í ÇÔ²² »ç¿ëÇÒ ¼ö ÀÖ´Â 7°¡Áö vector°¡ º°µµ ÆÇ¸ÅµÇ°í ÀÖ´Ù.

SMART library´Â ±âÁ¸ÀÇ ÀüÅëÀûÀÎ ¹æ¹ý¿¡ ºñÇØ ¸Å¿ì ³ôÀº È¿À²·Î Àüü±æÀ̸¦ Æ÷ÇÔÇÏ´Â cloneÀ» Á¦ÀÛÇÒ ¼ö ÀÖ´Ù. µû¶ó¼­ SMART cDNA library¿¡¼­ ¼±º°µÈ cloneÀº mRNAÀÇ 5¡¢-untranslated regionÀ» ¿ÏÀüÈ÷ Æ÷ÇÔÇϰԵȴ٠(2).

RNA/cDNA Quality Assay¸¦ »ç¿ëÇÏ¸é ½ÇÇèÀ» ½ÃÀÛÇϱâ Àü¿¡ Ãʱ⠽÷á·Î »ç¿ëµÇ´Â human RNAÀÇ »óŸ¦ È®ÀÎÇÒ ¼ö ÀÖ´Ù. º» ½ÇÇè¿¡´Â RT-PCR¹ýÀÌ ÀÌ¿ëµÇ¹Ç·Î ½ÇÇè¿¡ Àû¿ëÇÒ full-length cDNA°¡ ÁõÆøµÇ´ÂÁö ¿©ºÎ¸¦ Á÷Á¢ÀûÀ¸·Î È®ÀÎÇÒ ¼ö ÀÖ´Ù. ±âÁ¸ÀÇ ½ÇÇè½Ç¿¡¼­ »ç¿ëÇÏ´ø ÀåºñµéÀ» ÀÌ¿ëÇϹǷΠºü¸£°Ô RNA »óŸ¦ È®ÀÎÇÒ ¼ö ÀÖÀ» »Ó¸¸ ¾Æ´Ï¶ó ºÒÆíÇϰí À¯ÇØÇÑ formaldehyde gelÀ» Á¦ÀÛÇÒ Çʿ䵵 ¾ø´Ù.


Figure 1. Comparison of SMART cDNA synthesis vs. conventional synthesis for library construction.

Á¦Ç°¸í Cat. No. ¿ë·® ¸Å´º¾ó First Strand Synthesis cDNA Amplification Vector Ư¡
SMART cDNA
Library Construction
Kit
634901 10 ȸ ¸Å´º¾ó SMART IV Oligonucleotide 5' PCR Primer ¥ëTriplEx2
(lamda phage)
* SMART cDNA ÇÕ¼º ±â¼ú
* lamda phage¿¡ library Á¦ÀÛ
* Á¦ÀÛµÈ library¸¦ °í°´ÀÌ ¿øÇÏ´Â plasmid·Î coversion±ÇÀåÇÏÁö ¾ÊÀ½
CDS III/3' PCR Primer
In-Fusion SMARTer
Directional
cDNA Library
Construction Kit
634933 10 ȸ ¸Å´º¾ó SMARTer V Oligonucleotide 5' PCR Primer II A pSMART2IFD
Linearized
Vector
* SMART񃬣 In-Fusion Cloning 񃬣
* ¿øÇÏ´Â vector¿¡ °ð¹Ù·Î library
Á¦ÀÛ °¡´É(È¿À² °í·Á ÇÊ¿ä)
* InsertÀÇ cloning ¹æÇ⼺ êó
3¡¯ In-Fusion SMARTer CDS Primer 3¡¯ In-Fusion SMARTer PCR Primer


Keyword :

SMART¢ç Full length cDNA ÇÕ¼º ¡¤ Library Á¦ÀÛ
>
SMART¹ýÀ» ÀÌ¿ëÇÑ new version 5¡¯-/3¡¯-RACE K..
RACE 5/3 Kit (Unknown region analysis)
>
SMART¹ýÀ» ÀÌ¿ëÇÑ Full-length cDNA ÇÕ¼º
Full length cDNA Synthesis Kit
>
5¡¯-RACE¿Í 3¡¯-RACE·Î ºÎÅÍ cDNAÀ» È®º¸
Marathon cDNA Amplification Kit
>
È®½ÇÇϰí È¿°úÀûÀÎ cDNA Library ±¸Ãà
In-Fusion¢ç SMARTer cDNA Library Construction Kit