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Library screening, cloning and sequencing
Two degenerate primers (TL3, 5¡é-TTYGAICAYCCITAYCCICCIACIAARYIIAGTTT-
3¡é and TL4, 5¡é-CATIGGGTCRATICCRTTIGGICCIGCIACIGTNGG-
3¡é) were designed from AtTTG1 to amplify 700 bp from putative cotton homologues.Amplified fragments were ligated to pGEM-T Easy (Promega) and used to transform E. coli DH5-a F0 cells (Promega). The insert from one clone was released by EcoRI digestion, oligolabelled (Hodgson and Fisk, 1987) and used to
probe a 12 DPA (days post-anthesis) cotton fibre cDNA library.

resultÖÐÓëÖ®ºôÓ¦µÄ½á¹û£º
Initially, two degenerate TTG1 PCR primers were used to amplify a 700 bp fragment from cotton genomi cDNA. Nucleotide sequencing of four clones
revealed two distinct, but very similar, sequences.

ÎÊÌ⣺×îºóÔõô»á³öÏÖ Nucleotide sequencing of four clones£¿Õâ¸öfourÊÇÔõôÀ´µÄ£¿ÓÃ1¶Ô¼ò²¢ÒýÎïÈ¥À©Ôö700bpµÄƬ¶Î£¬¼ÙÈç˵À©Ôö³öÁË4¸ö²»Í¬µÄƬ¶Î£¬Õâ4¸öƬ¶ÎÓÖÔõô·Ö¿ª£¿ÖصãÊÇÄǸö4 clonesÊÇÔõô»ØÊ£¿¿´ÁËÁ½ÆªÎÄÏ×¶¼ÓÐÕâ¸ö¡£
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hnndpt(½ð±Ò+5):·Ç³£¸Ðл£¡ 2010-10-07 17:03:56
ºÇºÇ£¬²»ÊÇÌáµ½Á¬½Óµ½ pGEM-T EasyÔØÌåÉÏÁËô£¬È»ºóÓÖËæ¼´²âÐòÁË¡£»òÕß¾­¹ýRFLPʹ֮·ÖÐÍ£¬×îºóÓÐÁ½¸öÃ÷ÏÔ²îÒìµÄpattern£¬ËÍÈ¥²âÐò£¬µÃµ½two distinct, but very similar, sequences¡£ÔÙ×Ðϸ¿´¿´ÎÄÕ£¬Ó¦¸ÃÓÐrandomly select »òÕßRFLP·ÖÐÍÖ®ÀàµÄ´ÊÓïµÄ¡£
The world is a fine place and worth fighting for. I agree with the second part.
2Â¥2010-10-07 08:43:54
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hnndpt

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Originally posted by brightfuture01 at 2010-10-07 08:43:54:
ºÇºÇ£¬²»ÊÇÌáµ½Á¬½Óµ½ pGEM-T EasyÔØÌåÉÏÁËô£¬È»ºóÓÖËæ¼´²âÐòÁË¡£»òÕß¾­¹ýRFLPʹ֮·ÖÐÍ£¬×îºóÓÐÁ½¸öÃ÷ÏÔ²îÒìµÄpattern£¬ËÍÈ¥²âÐò£¬µÃµ½two distinct, but very similar, sequences¡£ÔÙ×Ðϸ¿´¿´ÎÄÕ£¬Ó¦¸ÃÓÐrand ...

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3Â¥2010-10-07 17:03:42
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brightfuture01

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Originally posted by hnndpt at 2010-10-07 17:03:42:

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ÎÄÖÐûÓÐÌáµ½RFLP£¬Èç¹ûÓüò²¢ÒýÎïÀ©Ôö³öÀ´µÄÐòÁÐÁ¬½Óµ½ÔØÌåÒÔºó£¬Ôõô¶ÔÆä²âÐò£¿Ëæ»úÑ¡Ôñ¼¸¸ö¿Ë¡½øÐвâÐòÂð£¿ÕâÑùµÄ»°»á²»»á°ÑÀ©Ôö³öµÄ²»Í¬ÐòÁÐÒÅ©ÁË£¿£¨Æ©Èç˵À©Ôö³öÁË5¸ö²» ...

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ÎÒÔø¾­×ö¹ýÒ»¸öÀàËÆµÄ¹¤×÷£¬28¸ö¿Ë¡¾­¹ýRFLPºó·ÖΪÁË13¸öpattern£¬ËùÒÔÒªÊÇËÍÉÙÁ˵ϰ£¬¹À¼Æ¾Í²»»á³öÀ´Õâô¶àÁË¡£Ò»°ãÇé¿ö϶¼ÊÇ×öÒ»¸öRFLPµÄ£¬È»ºóËͲ»Í¬µÄPattern²âÐò¡£
The world is a fine place and worth fighting for. I agree with the second part.
4Â¥2010-10-07 19:01:14
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