Ingabe usasebenzisa i-Trizol ukuze ukhiphe i-RNA?
Gosh, ungangivumeli ngikhathazeke ngempilo yakho!
U-Trizol usezokhishwa inyumbazane, ungangitsheli ukuthi usasebumnyameni?

Ingxenye eyinhloko ye-Trizol reagent yi-phenol, futhi ama-organic reagent afana ne-chloroform nawo ayangezwa ekusebenzeni kokuhlola.Ukuchayeka isikhathi eside kuzodala umonakalo empilweni yakho, ikakhulukazi ukuthintana ngqo nesikhumba kungabangela ukusha kwamakhemikhali okubi kakhulu nezibazi ezingapheli.

Ngaphansi kwezimo ezijwayelekile, ozakwethu baselabhorethri banganciphisa kuphela izingozi ngokuzihlomisa ngokugcwele (ukugqoka amajazi elebhu, amagilavu ​​namamaski).

Futhi iseluleko sami kuwe sithi: ngicela ufake i-Trizol ngokuqondile ukuze ugweme ukulimala okuvela emthonjeni.

1. Umkhiqizo wokuqala wesizukulwane sokukhishwa kwe-RNA
Iphuma ngo-1987 (i-Trizol reagents)

inzuzo:
Ukukhipha okuningi okuphelele

Ububi:
Ama-reagents anobuthi njenge-guanidine isothiocyanate-phenol-chloroform ayadingeka;
Ukusebenza kanzima;
Udinga ukusebenzisa i-DNase ukuze ususe ukungcoliswa kwe-DNA

2.Umkhiqizo wesizukulwane sesibili wokukhishwa kwe-RNA
Phuma ngo-1994 (QIAGEN RNeasy kanye nemikhiqizo eminingi yasekhaya)

inzuzo:
Ikholomu indlela ye-centrifugal yamukelwa, okulula ukuyisebenzisa;

Ububi:
Kudingeka i-centrifuge ukususa udoti wezicubu
Udinga ukusebenzisa i-DNase ukuze ususe ukungcoliswa kwe-DNA

3.Umkhiqizo wesizukulwane sesithathu wokukhishwa kwe-RNA
Iphuma ngo-2005 (QIAGEN I-RNeasy Plus kanye no-ForegeneImikhiqizo yochungechunge lokuhlanza i-RNA)

inzuzo:
Sebenzisa ngokuqondile ikholomu yokususa i-DNA ukuze ususe i-DNA nemfucumfucu yezicubu ngesikhathi esisodwa, ukonga isikhathi nomzamo;
Kuphephile futhi kulula, asikho isidingo sokwengeza ikholomu yokuhlunga kanye ne-DNase ukuze ususe kalula ukungcola futhi ugweme ukungcoliswa kwe-DNA

Ake sibheke umehluko wokusebenza phakathi komkhiqizo wesizukulwane sokuqala (i-Trizol), umkhiqizo wesizukulwane sesibili (i-RNeasy) kanye nomkhiqizo wesizukulwane sesithathu (i-Forgene) wokukhishwa kwe-RNA:

Ukusebenza emfanekisweni kucacile: umkhiqizo wesizukulwane sesithathu unezinzuzo ezisobala

Amagama amathathu nje ongawachaza: kulula!Ngokushesha!Ukuphepha

Ishadi le-PK lemiphumela yokuhlolaokweisizukulwane sokuqala, isizukulwane sesibili kanye nemikhiqizo yesizukulwane sesithathu yokukhishwa kwe-RNA:

1. EI-lectrophoretogram

Isizukulwane sokuqala (1: Trizol)
Isizukulwane sesibili (2:QIAGEN I-RNeasy)
Isizukulwane sesithathu (3: QIAGEN I-RNeasy plus, 4: Foregene)

Hlobo luni lomkhiqizo omuhle, uzokwazi ngokuqhathanisa
I-RNA ikhishwe kusetshenziswa ikhithi ye-Foregene
Hhayi kuphela ukugxila okuphezulu kodwa futhi nobuqotho obuhle

2.qPCR igrafu (igrafu yejika yokukhulisa)

Isizukulwane sokuqala (i-Trizol)

Isizukulwane sesibili (sky bluekushoQIAGEN RNeasy-, ebomvukushoQIAGEN RNeasy +)

Isizukulwane sesithathu (okuluhlaza okwesibhakabhaka kusho Forenene, okuluhlaza kusho QIAGEN RNeasy plus)

(Qaphela: Isizukulwane sesibili se-QIAGEN RNeasy yegrafu ye-qPCR isho ukuthi ikhithi ayifaki i-DNase futhi ayisusi ukungcola kwe-DNA. Ngakho-ke, i-CT iphakeme futhi umphumela awunembile; I-QIAGEN RNeasy + isho ukuthi i-DNase yengeziwe futhi ukungcola kwe-DNA kususiwe, futhi umphumela unembile)

Noma ngabe i-electrophoretogram noma igrafu ye-PCR yokukhulisa ijika yokukhulisa amandla, ikhwalithi yemikhiqizo ye-Foregene ayikho ngaphansi kunemikhiqizo yangaphandle.