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wawa13998

新虫 (初入文坛)

[求助] 求高人指导设计简并引物

老板叫我研究一种新虫子(鳞翅目,非模式物种)的tan基因,要P全长,可我自己用CODEHOP设计的简并引物都木有全长
他叫我取编码区后对其,取一头一尾,这样设出来的引物我去网上检测了一下退火温度,结果是没有退火温度范围 求高人指导一下 这个全长的引物该怎么设
以下是我用来对比的序列

>gi|294459899|ref|NP_001170882.1| tan protein [Bombyx mori]
MRELNGTAVCEIQLGRRHAVPVIYVRGSHYEVGFDVGRTFSSIIKSFISNYANLRDFEREYKTDTGRNAY
DKTLANMEKRFPYYVKEIRGVADGANVPFYQLFLLQMDDLIGTINDNHVPRNDTGGCSSVAFKNPQHTIL
GHTEDAFTETLNHFYIMSAHIIPTPEDREHGAVEERFSSLCYAGHMPGYTMGYNENGMVFSINTLSPLLL
KPGNTPRTFITRKLLSAKSFPDAERILRDEGLGIGNGFSVNMIWTDTWGDTKLYNVEVAPDLKGDRSLVH
VQKYDQDTLVHCNIYNRINVIEVIGEIVGSSKSRLKAIHAHAAPKTRGDIAEILSDVTGKDFQVFSMQKD
AIIKTIAAGIFDVEKRTWSIYIDKPSISEPVAVLPIRFSELESNK

>gi|315493450|gb|ADU32897.1| tan [Heliconius melpomene malleti]
MKELNGTTVCEKYIGRRRAVPVLYLQGSHYEVGFDVGRNFANVIKSFLSSYANLRDFEKEYKTEVGRNAY
EKTLSNMKERFPYYVKEMQGVADGAEVPFHQLFLLQMDDIIGTINDNHIPRNDTGGCSSLAIKTPNSAVL
GHTEDAFSETLNHFYIMSAHIIPTDEDKERGAIEERFASLCYAGHLPGYTMGYNENGLVFSINTLSPLVL
KPGNTPRTFITRALLASKSYADAERILRDEGLGIANGFSINMIWTNKDGERKIYNAEVAPDLKGDRSIIN
IHNYVDEPLVHTNIYQRISIKEVIGPIIDSSTSRLKVIHGHPPPQSQNDVKDILSDTSGTDFQVFQEKKE
SIIKTIAAGLFELDKMTWSIFINKPNISEPVAVLPIRFSMLKAN

>gi|357603439|gb|EHJ63768.1| tan protein [Danaus plexippus]
MSFRTTWIFIFLTLGVLCEPNIGRRRAVPVLYLQGSHYEVGFDVGRNFASIIKSFLSSYENLRDFEKEYK
TITGQDAYEKTLANMKKRYPYYVKEMQGVADGANVPFHQLFLLQMDDLIGTINDNHIPRNDTGGCSSLAV
KTRKSAILAHTEDAFGETLNHFYIMSAHIIPDPEDKELGAVEERFASLCYAGHMPGYTMGYNENGLVFSI
NTLSPLELKPGNTPRTFITRALLASKNFSDAERILRDEGLGIGNGFSINMIWSDKCGDQKIYNVEVAPDL
KADRSVLNINNYVDEPLIHTNIYQRLSVKEVIGPIIDSSTARLKAIHAHPPPENGKDLKNILSDISGQDF
QVFQEKEDSIIKTIAAGVFELDKMTWSIYINKPNISEPVAVLPIRFSSLSKCES

>gi|296040353|dbj|BAJ07601.1| Tan [Papilio polytes]
MRELYGNSLSEKHIGRRRVVPVIYLQGSHYEVGYDVGRNFASVIQSFLKTYQGLKDFEREYNTAAGRAAY
DSTLVNMKQRYPYYVKEIQGVADGAKVPFYQLFLLHMDDIIGTINDKNSNRVDSGGCSDIAFKSPENVVL
GHTEDAPMETVNHFYIMSAHVIPTPEDRELGAVEERFASLCYAGQLPGYTMGFNEHGLVFTINTLSPLVL
KPGNTPRMFITRALLTARTPADAEKILLDEGLGIGNGFSTNVIWTYENGERDLMNIEVAPDLSANKSILN
IHKYTNETLMHCNAYLREKVQEVTGDIIDSSVTRMKAIHSHAPPRSRRDIESVLSDTTYKDYPVFQTRLD
APIQTIAAGIFDMSARTWSIYIDKPDSSEPVAILPIRFSQLNKNH

>gi|296040349|dbj|BAJ07599.1| Tan [Papilio machaon]
MRGLIGTSLSEKHIGRRRVVPVIYLQGSHYEVGYDVGRNFASIIQSFLKTYQGLKDFEREYNTPSGRAAY
DNTLLNMKQRYPYYVKEIQGVADGAKVPFYQLFLLHMDDIIGTINDKNSNRIDDGGCSDIAFKSPENVVL
GHTEDAPMETLNHFYIMSAHVIPTPEDRELGAVEERFASLCYAGQLPGYTMGFNEYGLVFTINTLSPLVL
KPGNTPRMFITRALLTARTPAEAEKILLDEGLGIGNGFSTNVIWTFENGERDLMNIEVAPDLTANKSILN
IQKYTNDTLMHCNAYLRERVQEVTGDIIESSVTRMKAIRRHAPPRSRHDIERVLSDTTYKDFPVFQTRLD
APIQTIAAGIFDMSAKTWSIYIDKPDSSEPVALLPMTFSQLNQNH

>gi|291486757|dbj|BAI87828.1| Tan [Papilio xuthus]
MRGLYGTSLSEKHIGRRRVVPVIYLQGSHYEVGYDVGRNFASIIQSFLKTYQGLKDFEREYNTPSGRAAY
DNTLLNMKQRYPYYVKEIQGIADGAKVPFYQLFLLHMDDIIGTINDKTSNRIDDGGCSDIAFKSPENVVL
GHTEDAPMETVNHFYIMSAHVIPTPEDRELGAVEERFASVCYAGQLPGYTMGFNEYGLVFTINTLSPLLL
KPGNTPRMFITRALLTARTPAEAEKILLDEGLGIGNGFSTNVIWTSENGERDLMNIEVAPNLSGNKSILN
IQKYTNDTLMHCNAYLRERVQEVTGDIIESSVTRMKAIRRHSPPRSRRDIERVLSDTTYKDFPVFQTRLD
APIQTIAAGIFDMSAKTWSIYIDKPDSSEPVAILPIRFSQLNQNHHN
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