Attached is the sequence file of my gene and my target region is in first 400bp. Many thanks for consideration.
LAMP primers design result is attached
alternative sets with FastPCR:
PrimerID Sequence(5'-3') Tm(°C) Primer_Quality(%) Fragment_Size(bp)/Tm(°C) Topt(°C)
F3_:1F3_1_56-76 ttgcgaccaccgtacaagtgc 60.0 85
F2_:1F5_1_92-116 aaacatgaacacttccgttggacta 56.4 88
F1_:1F8_1_131-155 cttcgtctgttcacagttcatccga 58.8 81
B1_:1R15_1_162-183 gctcctcgtcttctgaacacca 58.3 82
B2_:1R11_1_218-242 catgttctggactggcctgattagt 58.3 78
B3_:1R8_1_243-267 caaactggacattgactttttcggt 56.5 80
FIP tcggatgaactgtgaacagacgaag-aaacatgaacacttccgttggacta 69.8
BIP tggtgttcagaagacgaggagc-catgttctggactggcctgattagt 70.4 212
B3_:1R7_1_258-282 gaacgaatgctaaaccaaactggac 57.2 80
BIP tggtgttcagaagacgaggagc-catgttctggactggcctgattagt 70.4 227
B2_:1R10_1_222-244 gtcatgttctggactggcctgat 58.4 75
BIP tggtgttcagaagacgaggagc-gtcatgttctggactggcctgat 70.4 227
B1_:1R14_1_167-188 cagacgctcctcgtcttctgaa 58.1 80
BIP ttcagaagacgaggagcgtctg-catgttctggactggcctgattagt 70.2 212
BIP ttcagaagacgaggagcgtctg-catgttctggactggcctgattagt 70.2 227
BIP ttcagaagacgaggagcgtctg-gtcatgttctggactggcctgat 70.2 227
B1_:1R13_1_177-201 aaagatctctaaccagacgctcctc 57.7 81
BIP gaggagcgtctggttagagatcttt-catgttctggactggcctgattagt 68.9 212
BIP gaggagcgtctggttagagatcttt-catgttctggactggcctgattagt 68.9 227
BIP gaggagcgtctggttagagatcttt-gtcatgttctggactggcctgat 68.9 227
B2_:1R9_1_232-256 ttgactttttcggtcatgttctgga 56.6 80
BIP gaggagcgtctggttagagatcttt-ttgactttttcggtcatgttctgga 68.9 227
B1_:1R12_1_181-205 ctgaaaagatctctaaccagacgct 56.4 86
BIP agcgtctggttagagatcttttcag-catgttctggactggcctgattagt 68.7 212
FastPCR alternative sets with Loop primers
Thank you very much respected Ruslan Kalendar , for your kind help.
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