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zxhong

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[交流] 【原创】求昆虫 COI引物设计或选用


我想用COI引物,但在NCBI上查到同一科的昆虫用的COI引物不同 ,
1.有的用两个引物lLCO和HCO及C1-J-2183和 TL2-N-3014,
LCO,      5′-GGT CAA CAA ATC ATA AAG ATA TTG G-3 ′;
HCO,      5′-TAA ACT TCA GGG TGA CCA AAA AAT CA-3′
C1-J-2183,  5′-CAA CAT TTA TTT TGA TTT TTT GG-3′;
TL2-N-3014, 5′-TCCAATGCACTAATCTGCCATATTA-3′
2.有的用其中一个:
LCO, 5′-GGT CAA CAA ATC ATA AAG ATA TTG G-3 ′;
HCO, 5′-TAA ACT TCA GGG TGA CCA AAA AAT CA-3′
3.有的上游用C1-J-2183,下游自己设计:
(C1-J-2183)(5′ CAA CAT TTA TTT TGA TTT TTT GG 3′) (reproduced from Simon et al.,1994), and a modified version of C1-N-2968 (Sperling et al., 1997) named K866(C1-N-2972) (5′GTA TTT CGT TAT AA/T/C/GG AA/GT GTT 3′). Becausethis sequence was too long to be sequenced in one step, two internal primers weredesigned for amplification with the end-primers, namely LimH2929
(C1-N-2609)(5′CGA ATA CTG CTC CTA TTG ATA 3′) to work with Jerry, and Nils (C1-J-2387) (5′TCA CCA TCA ATA TTG TGA AC 3′) to work with K866.
4.有的用两对引物为:
C1-J-2183 Forward 5_- CAACATTTATTTTGATTTTTTGG-3_
TL2-N-3014 Reverse 5_-TCCAATGCACTAATCTGCCATATTA-3_
C1-J-2530 Forward 5_-GGAGTAATTCTAGCCAACTC-3_
C1-N-2609 Reverse 5_- GAATACTGCTCCTATGGATA-3_
5.有的用:
LR-J-13417 (5‘-CGCCTGTTTAACAAAAACAT-3‘) (Simon et al., 1994)
LRJ-12961 (5‘-TTTAATCCAACATCGAGG-3’) (Cognatoand Vogler, 2001).

两个引物即两个反应,测序费用太高(一个物种测两个反应,两份费用),我想用一个引物完成实验。我可能用他们的部分结果比对,我该如何选用引物?上面哪些引物即经济、结果又能互相匹配(结果互相比对)。
不同的引物,产物可否互相比对。
请高手指点,多谢 !
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