Kodi mukugwiritsabe ntchito Trizol kuchotsa RNA?
Gosh, musandilole ine nkhawa za thanzi lanu!
Trizol watsala pang'ono kutsanulidwa, osandiuza kuti ukadali mumdima?

Chigawo chachikulu cha Trizol reagent ndi phenol, ndi organic reagents monga chloroform nawo anawonjezera mu ntchito experimental.Kuwonekera kwa nthawi yayitali kungawononge thanzi lanu, makamaka kukhudzana kwachindunji pakhungu kungayambitse kutentha kwakukulu kwa mankhwala ndi zipsera zosatha.

Nthawi zonse, ogwira nawo ntchito mu labotale amatha kuchepetsa zoopsazo podzikonzekeretsa (kuvala malaya a labu, magolovesi ndi masks).

Ndipo upangiri wanga kwa inu ndikuti: chonde sinthani Trizol mwachindunji kuti mupewe kuvulaza komwe kumachokera.

1. M'badwo woyamba mankhwala a RNA m'zigawo
Anatuluka mu 1987 (Trizol reagents)

ubwino:
More wathunthu m'zigawo

Zoyipa:
Ma reagents oopsa monga guanidine isothiocyanate-phenol-chloroform amafunikira;
ntchito zovuta;
Muyenera kugwiritsa ntchito DNase kuchotsa kuipitsidwa kwa DNA

2.Chinthu chachiwiri cha m'badwo wa RNA
Tulukani mu 1994 (QIAGEN RNeasy ndi zinthu zambiri zapakhomo)

ubwino:
Column centrifugal njira imatengedwa, yomwe ndi yosavuta kugwiritsa ntchito;

Kuipa:
M'pofunika centrifuge kuchotsa zinyalala minofu
Muyenera kugwiritsa ntchito DNase kuchotsa kuipitsidwa kwa DNA

3.M'badwo wachitatu mankhwala a RNA m'zigawo
Ikutuluka mu 2005 (QIAGEN RNeasy Plus ndi ForegeneZogulitsa za RNA purification series)

ubwino:
Gwiritsani ntchito mzere wochotsa DNA mwachindunji kuchotsa DNA ndi zinyalala za minofu nthawi imodzi, kupulumutsa nthawi ndi khama;
Zotetezeka komanso zosavuta, palibe chifukwa chowonjezera zosefera ndi DNase kuti muchotse zonyansa ndikupewa kuipitsidwa kwa DNA.

Tiyeni tiwone kusiyana kwa magwiridwe antchito pakati pa chinthu cham'badwo woyamba (Trizol), cham'badwo wachiwiri (RNeasy) ndi cham'badwo wachitatu (Forgene) cha RNA:

Kugwira ntchito pachithunzichi kumamveka bwino: mankhwala a m'badwo wachitatu ali ndi ubwino woonekeratu

Mawu atatu okha oti afotokoze: zosavuta!Mofulumira!Chitetezo

Tchati cha PK chazotsatira zoyesererazam'badwo woyamba, wachiwiri ndi wachitatu wa RNA m'zigawo:

1. Electrophoretogram

M'badwo woyamba (1: Trizol)
M’badwo wachiwiri (2:QIAGEN RNeasy)
M’badwo wachitatu (3: QIAGEN RNeasy kuphatikiza, 4: Foregene)

Ndi mtundu wanji wa mankhwala omwe ali abwino, mudzadziwa poyerekeza
RNA yotengedwa pogwiritsa ntchito zida za Foregene
Osati kungoganizira kwambiri komanso kukhulupirika kwabwino

2.qPCR graph (kukulitsa curve graph)

M'badwo Woyamba (Trizol)

M'badwo wachiwiri (sky bluekutanthauzaQIAGEN RNeasy-, wofiirakutanthauzaQIAGEN RNeasy +)

M'badwo wachitatu (buluu amatanthauza Foregene, wobiriwira amatanthauza QIAGEN RNeasy plus)

(Zindikirani: Mbadwo wachiwiri wa QIAGEN RNeasy wa qPCR graph umatanthauza kuti chida sichikuwonjezera DNase ndipo sichichotsa kuipitsidwa kwa DNA. Choncho, CT ndi yapamwamba ndipo zotsatira zake sizolondola; QIAGEN RNeasy + imatanthauza kuti DNase yawonjezedwa ndipo kuipitsidwa kwa DNA kwachotsedwa, ndipo zotsatira zake ndi zolondola)

Kaya ndi electrophoretogram kapena kuchuluka kwa PCR amplification curve graph, mtundu wa zinthu za Foregene sizotsika poyerekeza ndi zomwe zatumizidwa kunja.