RT-LAMP agba agba (bọọlụ Lyophilized)
Ngwaahịa a nwere ihe nchekwa mmeghachi omume, RT-Enzymes Mix (Bst DNA polymerase na reverse transcriptase okpomọkụ na-eguzogide), ihe nchebe lyophilized na ihe ndị na-esiji agba chromogenic.Ngwaahịa bụ ụdị bọọlụ lyophilized, na-eji naanị primers na ndebiri.Ngwa a na-enye ngwa ngwa, nchọpụta anya doro anya nke mmụba, nke mmeghachi omume na-adịghị mma na-egosi na-acha uhie uhie na mmeghachi omume dị mma na-egosipụta site na mgbanwe na-acha odo odo.
Akụkụ
Ngwakọta agba agba RT-LAMP (Beads Lyophilized)
Ngwa
Maka DNA ma ọ bụ RNA isothermal amplification.
Ọnọdụ Nchekwa
A na-ebugharị ma chekwaa ya na 2 ~ 8 ℃.Ngwaahịa a dị irè maka ọnwa iri na abụọ.
Protocol
1.Wepụta nọmba kwekọrọ na ntụ ntụ Lyophilized dị ka ọnụọgụ ule si dị.
2.Kwadebe ngwakọta mmeghachi omume
| Akụkụ | Olu |
| Ngwakọta agba agba RT-LAMP (Beads Lyophilized) | 1 iberibe (2 beads) |
| 10 × Ngwakọta mbụa | 5 μl |
| Ụdị DNA/RNA b | 45 μl |
ndetu:
1. 10 × Primer Mix Concentration: 16 μM FIP / BIP, 2 μM F3 / B3, 4 μM Loop F / B;
2. A na-atụ aro ka agbaze ndebiri acid nucleic site na iji mmiri DEPC.
3.Tinye na 65 Celsius C maka 30-45mins, nke enwere ike ịgbatị nke ọma dịka oge mgbanwe agba si dị.
4.Dị ka anya gba ọtọ si kwuo, odo na-adị mma na uhie dị njọ.
Ihe ndetu
1.Enwere ike ịhazi okpomọkụ mmeghachi omume n'etiti 62 ℃ na 68 ℃ dị ka ọnọdụ primer si dị.
2.The ngwugwu reagents ekwesịghị ikpughe na ikuku ruo ogologo oge.
3.Mmeghachi omume ngbanwe nke uhie na odo na-adabere na pH mgbanwe nke sistemu mmeghachi omume, biko ejila ihe nchekwa nchekwa Tris nucleic acid nwere, akwadoro iji ddH2O echekwabara nucleic acid.
4.A ga-eme nnwale ahụ n'ụzọ ziri ezi, gụnyere nkwadebe nke usoro mmeghachi omume, ọgwụgwọ nlele na mgbakwunye ihe atụ.
5.A na-atụ aro ka ịkwadebe usoro mmeghachi omume na tebụl dị ọcha ma tinye ndebiri na mkpuchi uzuzu nke ụlọ ndị ọzọ iji zere ụgha.
