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little7tone

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[求助] 引物设计问题,各位大神帮帮忙!! 已有1人参与

小女子走上一条类似转专业的性质。第一次做PCR,好多不懂,各位大神帮帮忙!!!
要设计鉴定副溶血性弧菌的引物
1、我把目的基因定为它的TLH基因,可是NCBI里面有有关的TLH基因(product="thermolabile),还有TL基因(product="thermolabile hemolysin),有没有大神知道他们是不是一样的?
2、怎么找目的基因高度保守区?都说是序列比对,怎么比对?怎么blast?
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snoopytbw

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【答案】应助回帖

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little7tone(西门吹雪170代发): 金币+2, 鼓励回帖交流 2014-08-19 20:38:20
little7tone: 金币+1, ★有帮助 2014-08-21 09:46:49
找到几个可能基因
VS_II0201 Thermolabile hemolysin (TL)-(Lecithin-dependent hemolysin) (LDH) [ Vibrio splendidus LGP32 ]
但是也不是副溶血,是灿烂弧菌
NCBI Reference Sequence: NC_011744.2

Vibrio parahaemolyticus strain C1 Tlh (tlh) gene, partial cds
GenBank: JQ929914.1
http://www.ncbi.nlm.nih.gov/nucc ... amp;amp;strand=true

Vibrio parahaemolyticus strain CECT 5184 thermolabile hemolysin (tl) gene, partial cds
GenBank: EU546143.1
http://www.ncbi.nlm.nih.gov/nuccore/189031200?report=fasta

知道跟你说的一样么。你在核苷酸(nucleotide)里面找,直接搜 TL或者TLH Vibrio parahaemolyticus
2楼2014-08-19 13:49:35
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little7tone

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引用回帖:
2楼: Originally posted by snoopytbw at 2014-08-19 13:49:35
找到几个可能基因
VS_II0201 Thermolabile hemolysin (TL)-(Lecithin-dependent hemolysin) (LDH)
但是也不是副溶血,是灿烂弧菌
NCBI Reference Sequence: NC_011744.2

Vibrio parahaemolyticus strain C1  ...

不懂,我是用“TLH Vibrio parahaemolyticus”搜出来是48个,用“TL Vibrio parahaemolyticus”搜出了是13个,这两个基因是一样的吗?
3楼2014-08-19 19:17:20
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