TaKaRa MiniBEST Plant RNA Extraction Kit´Â ½Ä¹°Á¶Á÷ ¹× ¼¼Æ÷·ÎºÎÅÍ °í¼øµµÀÇ RNA¸¦ ºü¸£°Ô ÃßÃâÇÒ ¼ö ÀÖ´Â small-scaleÀÇ RNA extraction Á¦Ç°ÀÌ´Ù.
RNA¸¦ ºü¸£°í Æí¸®ÇÏ°Ô ÃßÃâÇϱâ À§Çؼ »ùÇÿ¡ µû¶ó ÃÖÀûÈµÈ 2Á¾·ùÀÇ lysis buffer¸¦ »ç¿ëÇÑ´Ù. ÀϹÝÀûÀÎ ½Ä¹°Á¶Á÷ (ÀÙ, ÁÙ±â, À¯¹¦ µî) ¶Ç´Â RNA ÃßÃâÀÌ
¾î·Á¿î polysaccharide, polyphenolÀ» ´Ù·® ÇÔÀ¯ÇÑ ½Ä¹°Á¶Á÷ (°ú½Ç, Á¾ÀÚ µî)À¸·ÎºÎÅÍ 200 bp ÀÌ»óÀÇ RNA¸¦ °í¼øµµ·Î ÃßÃâÇÒ ¼ö ÀÖ´Ù (200 bp ÀÌÇÏÀÇ small RNAÀÇ Á¤Á¦µµ °¡´ÉÇϸç,
À̸¦ À§ÇÑ protocolÀº º°µµ ÷ºÎµÇ¾î ÀÖ´Ù).
º» Á¦Ç°À» »ç¿ëÇÏ¸é ¼¼Æ÷³ª Á¶Á÷ÀÇ lysis, ¶Ç´Â ±ÕÁúÈ (homogenization) ÈÄ, phenolÀ̳ª chloroformÀÇ À¯±â¿ë¸Å »ç¿ë¾øÀÌ 30ºÐ À̳»¿¡ ´Ü¹éÁú°ú gDNAÀÇ ¿À¿° ¾ø´Â °í¼øµµÀÇ RNA¸¦ ÃßÃâÇÒ ¼ö ÀÖ´Ù.
Á¤Á¦µÈ RNA´Â northern blotting, dot blotting, mRNA purification, in vitro translation, RNase protection assay, RT-PCR, Real Time PCR, cDNA library construction°ú
°°Àº ´Ù¾çÇÑ ºÐÀÚ»ý¹°ÇÐ ½ÇÇè¿¡ Àû¿ë °¡´ÉÇÏ´Ù.
±×¸² 1. TaKaRa MiniBEST Plant RNA Extraction KitÀÇ workflow
M : DL2,000 DNA Marker
1 : ¼ö¼ö À¯¹¦
2 : ¿Á¼ö¼ö À¯¹¦
3 : ³ìµÎ À¯¹¦
4 : Çعٶó±â À¯¹¦
±×¸² 2. À¯¹¦¿¡¼ Total RNAÃßÃâ. TaKaRa MiniBEST Plant RNA Extraction KitÀÇ Protocol-IÀ» »ç¿ëÇÏ¿©, 100 §·ÀÇ ´Ù¾çÇÑ ½Ä¹° À¯¹¦ (seedling)·ÎºÎÅÍ
total RNA¸¦ ÃßÃâÇÏ¿´´Ù. ÃßÃâµÈ RNAÀÇ Àü±â¿µµ¿ °á°ú, °í¼øµµÀÇ RNA°¡ ÃßÃâµÈ °ÍÀ» È®ÀÎÇÏ¿´´Ù
M : DL2,000 DNA Marker
1 : ¶¥Äá Á¾ÀÚ
2 : Çعٶó±â Á¾ÀÚ
3 : ¿Á¼ö¼ö Á¾ÀÚ
±×¸² 3. ½Ä¹°ÀÇ Á¾ÀÚ¿¡¼ Total RNAÃßÃâ. TaKaRa MiniBEST Plant RNA Extraction KitÀÇ Protocol-II¸¦ »ç¿ëÇÏ¿©, 50 §·ÀÇ
´Ù¾çÇÑ ½Ä¹° Á¾ÀڷκÎÅÍ total RNA¸¦ ÃßÃâÇÏ¿´´Ù. ÃßÃâµÈ RNAÀÇ Àü±â¿µµ¿ °á°ú, °í¼øµµÀÇ RNA°¡ ÃßÃâµÈ °ÍÀ» È®ÀÎÇÏ¿´´Ù
M : DL2,000 DNA Marker
1 : ¹è °ú½Ç
2 : »ç°ú °ú½Ç
3 : Å丶Åä °ú½Ç
±×¸² 4. ½Ä¹°ÀÇ °ú½Ç¿¡¼ Total RNAÃßÃâ. TaKaRa MiniBEST Plant RNA Extraction KitÀÇ Protocol-I¸¦ »ç¿ëÇÏ¿©, 100 §·ÀÇ ´Ù¾çÇÑ ½Ä¹° °ú½Ç·ÎºÎÅÍ total RNA¸¦
ÃßÃâÇÏ¿´´Ù. ÃßÃâµÈ RNAÀÇ Àü±â¿µµ¿ °á°ú, °í¼øµµÀÇ RNA°¡ ÃßÃâµÈ °ÍÀ» È®ÀÎÇÏ¿´´Ù