- ºü¸¥ genomic DNAÀÇ PCR-based walking
- Exon-intron junctions, promoters, ¶Ç´Â ´Ù¸¥ regulatory element¸¦ Ž»öÇϴµ¥ ÀûÇÕ
- ¹ÌÁöÀÇ ¼¿À» ½Å¼ÓÇÏ°Ô È®ÀÎ
GenomeWalker Kit´Â expressed sequence tag (EST)°ú °°ÀÌ ÀÌ¹Ì ¾Ë°í ÀÖ´Â genomic DNA ¼¿·ÎºÎÅÍ PCRÀ» ÀÌ¿ëÇÏ¿© ¹ÌÁöÀÇ ¼¿À» È®ÀÎÇϱâ À§ÇÑ Á¦Ç°ÀÌ´Ù.
GenomeWalker´Â ±ä ¼¿À» ÁõÆøÇÏ´Â PCR ±â¹ý (long-distance PCR)À» ÀÌ¿ëÇÏ¿© °¢ ´Ü°è¸¶´Ù genomic DNA 6 kb ÀÌ»óÀ» È®ÀÎÇÒ ¼ö ÀÖ´Ù. WalkingÀº ÀÌÀü ´Ü°è¿¡¼
¾òÀº ¿°±â¼¿À» ±âº»À¸·Î »õ·Î¿î primer¸¦ Á¦ÀÛÇÏ¿© WalkingÀ» ¹Ýº¹ÇÏ´Â °Í¸¸À¸·Î °£´ÜÇÏ°Ô ¹üÀ§¸¦ ³ÐÈú ¼ö ÀÖ´Ù.
GenomeWalker Universal Kit
GenomeWalker Universal Kit´Â Á¾¿¡ »ó°ü¾øÀÌ ±× Á¾ÀÇ genomeÀ¸·ÎºÎÅÍ GenomeWalker library¸¦ Á¦ÀÛÇÒ ¼ö ÀÖ´Ù. ÀÌ Universal Kit´Â °¢±â ´Ù¸¥ 3Á¾ÀÇ ½Ã·á·ÎºÎÅÍ
library¸¦ Á¦ÀÛÇÏ¿© 80ȸÀÇ walk¸¦ ÁøÇà ÇÒ ¼ö ÀÖ´Ù.
Kit¸¦ ÀÌ¿ëÇÒ ¶§´Â °¢°¢ÀÇ À¯ÀüÀÚ¿¡ ¸Â°Ô gene-specific primer (GSP)¸¦ Á¦ÀÛÇØ¾ß Çϸç, long-distanc PCR (LD-PCR)¿¡ ÀûÇÕÇÑ DNA polymerase¸¦ º°µµ·Î ±¸¸ÅÇØ¾ß ÇÑ´Ù.
Clontech¿¡¼´Â Advantage 2 Polymerase Mix ¶Ç´Â Advantage 2 PCR Kit¸¦ ÃßõÇÑ´Ù.
Universal GenomeWalker 2.0
Universal GenomeWalker 2.0Àº Á¾¿¡ »ó°ü¾øÀÌ ±× Á¾ÀÇ genomic DNAÀ¸·ÎºÎÅÍ GenomeWalker library¸¦ Á¦ÀÛ¿¡ ÇÊ¿äÇÑ ¸ðµç ½Ã¾àÀÌ Æ÷ÇԵǾî ÀÖ´Ù.
ÀÌ Å°Æ®¿¡´Â
library¸¦ ±¸ÃàÇÒ ¼ö ÀÖ´Â component (PCR polymerase Æ÷ÇÔ), genomic DNA Á¤Á¦ kit, digested DNA clean-up kit°¡ Æ÷ÇԵǾî ÀÖ´Ù.
ÀÌ Universal Kit´Â °¢±â
´Ù¸¥ 3Á¾ÀÇ ½Ã·á·ÎºÎÅÍ library¸¦ Á¦ÀÛÇÏ¿© 80ȸÀÇ walk¸¦ ÁøÇà ÇÒ ¼ö ÀÖ´Ù.
Kit¸¦ ÀÌ¿ëÇÒ ¶§´Â °¢°¢ÀÇ À¯ÀüÀÚ¿¡ ¸Â°Ô gene-specific primer (GSP)¸¦ Á¦ÀÛÇØ¾ß ÇÑ´Ù.

Figure 1. The GenomeWalker method.
Components & Storage Conditions
°¢ Á¦Ç° ±¸¼º¹°°ú º¸°üÁ¶°ÇÀº Certificate of Analysis ¸¦ ÂüÁ¶ÇϽʽÿÀ.