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Ášnµ„´ 25 ôl Ÿ¤Ä®oÁ oµ„´œ n¤š¸É°»–®£¼¤· 42 OC Áž}œÁª¨µ 1 ´ÉªÃ¤Š ‹µ„œ´ÊœªµŠÅªoĜœÊε Ȋ¨³Á˜·¤ DEPC treated
H2O ž¦· ¤µ˜¦ 55 ôl ‹³Å—ož¦·¤µ˜¦¦ª¤ 80 ôl œÎµ sscDNA °Š˜´ª°¥nµŠ˜nµŠÇ ŞÁ¡·É ¤ž¦·¤µ– cDNA °Š
TDFREP241 ¨³¥¸œ 18S rRNA Ž¹ÉŠÁž}œ internal control ĜÁ‡¦ºÉ °Š PCR (iCycler iQTM :Biorad ) ˜´ª°¥nµŠ¨³ 3 ŽÊε
×¥ž¦´˜´ÊŠÃž¦Â„¦¤„µ¦šÎ µŠµœ—´Šœ¸Ê °»–®£¼¤· 94 oC Áª¨µ 2 œµš¸ ‹Îµœªœ 1 ¦° °» –®£¼¤· 94 oC Áª¨µ 30 ª·œµš¸
°»–®£¼¤· 56 oC Áª¨µ 30 ª·œµš¸ ¨³°»–®£¼¤· 72 oC Áª¨µ 45 ª· œµš¸ ‹Îµœªœ 30 ¦° 宦´  TDFREP241 ¨³‹Îµœªœ
15 ¦°Îµ®¦´ ¥¸œ 18S rRNA œÎ µ PCR product ޚε°·Á¨È„ڦ⦸Ž· ĜÁ‹¨°³„µÃ¦‡ªµ¤Á o¤ oœ 2% ¥o°¤Á‹¨
—oª¥ ethidium bromide œÎµÅž™nµ¥¦¼ž ¨³ª´—‡nµ intensity °ŠÂ™ cDNA °Šš»„˜´ ª°¥nµŠ 宦´ ‡nµ„µ¦Â—Š°°„
°Š¥¸œ °ŠÂ˜n¨³˜´ª°¥nµŠ‡Îµœª–‹µ„‡nµ°´˜¦µnªœ¦³®ªnµŠ intensity °ŠÂ™ cDNA °Š TDFREP241 ˜n° intensity
°ŠÂ™ cDNA °Š¥¸œ 18S rRNA
ª·Á‡¦µ³®ro°¤¼¨šµŠ™·˜·
ª·Á‡¦µ³®ro °¤¼¨Ÿ¨ °ŠÃŽÁ—¸ ¥¤‡¨°Å¦—r ᨸÁ°›¸ ¨¸œÅ„¨‡°¨Â¨³„µ¦„¦¸—˜n °„µ¦Â—Š°°„ °Š
TDFREP241 °Š˜´ª°¥nµŠ˜nµŠÇ ×¥„µ¦š—°‡n µ t (t-test) nªœ„µ¦Â—Š°°„ °Š TDFREP241 Ĝ„¨oµ¥µŠ„¨»n ¤
Ä®o Ÿ¨Ÿ¨·˜œÊÎ µ¥µŠ¼ŠÂ¨³˜Éεŗoš—°‡ªµ¤Âž¦ž¦ªœ (ANOVA) ¨³š—°‡ªµ¤Â˜„˜nµŠ °Š‡n µÁŒ¨¸É ¥Ã—¥ª·›¸
Duncan‘s multiple range test (DMRT)
Ÿ¨„µ¦ª·‹´ ¥Â¨³°£·ž¦µ¥Ÿ¨
„µ¦Â—Š°°„ °Š TDFREP241 ÁœºÊ°Á¥ºÉ°˜nµŠÇ °Š˜oœ„¨oµ¥µŠ¡µ¦µ
‹µ„„µ¦ª·Á‡¦µ³®r „µ¦Â—Š°°„ °Š TDFREP241 ĜÁœºÊ°Á¥ºÉ°Ä „oµœÄ ¨Îµ˜oœÂ¨³¦µ„ °Š˜oœ„¨oµ¥µŠ¡µ¦µ
µ¥¡´œ›»r RRIM600 °µ¥» 3 Á—º°œ ¡ªnµ¤¸‡ªµ¤Â˜„˜nµŠ„´œ°¥nµŠ¤¸ œ´¥Î µ‡´šµŠ™· ˜·š¸É ¦³—´ 0.05 ץĜÁœºÊ °Á¥ºÉ°¨Îµ
˜oœ¤¸ „µ¦Â—Š°°„ °Š¥¸œ¤µ„š¸É »— ¦°Š¨Š¤µ‡º° „o µœ ¨³¦µ„ nªœÄœÁœºÊ°Á¥ºÉ°Ä‹³¤¸„µ¦Â—Š°°„ °Š¥¸œœo°¥
š¸É »— (£µ¡š¸É 1) „µ¦Â—Š°°„ °Š TDFREP241 °—‡¨o°Š„´ž¦·¤µ– °Š laticifer cell ĜÁœºÊ°Á¥ºÉ°—´Š„¨nµª Ž¹ÉŠ
ÁœºÊ°Á¥ºÉ° °Š„¨oµ¥µŠ¡µ¦µ‹³¤¸ laticifer cell ¤µ„š¸É »—ÄœÁœºÊ°Á¥ºÉ°¨Îµ˜oœÂ¨³œo°¥š¸É »—ÄœÁœºÊ°Á¥ºÉ°Ä (Priya, 2007)
£µ¡š¸É 1 „µ¦šÎ µ°·Á¨‡Ãš¦Ã¢¦¸Ž· RT-PCR product °Š TDFREP241 ¨³¥¸ œ 18S rRNA ĜÁ‹¨°³„µÃ¦‡ªµ¤
Á o¤ oœ 2 Áž°¦r ÁŽÈœ˜r („) ¨³‡nµÁŒ¨¸É¥„µ¦Â—Š°°„ °Š TDFREP241 ( ) ‹µ„ÁœºÊ°Á¥ºÉ°Ä „oµœÄ ¨Îµ˜oœ ¨³¦µ„ °Š
„¨oµ¥µŠ¡µ¦µµ¥¡´œ›»r RRIM600
ĝ „oµœÄ ¨Îµ˜oœ ¦µ„
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ÁœºÊ°Á¥ºÉ °˜nµŠÇ
Relative expression levels
TDFREP241
18S rRNA
ĝ „o µœÄ ¨Îµ˜oœ ¦µ„
c
b
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