Uchiri kushandisa Trizol kubvisa RNA?
Gosh, usandirega ndichinetseka nezvehutano hwako!
Trizol yave kuda kuiswa marginalized,usandiudze kuti uchiri murima here?

Chinhu chikuru cheTrizol reagent ndeye phenol, uye organic reagents senge chloroform inowedzerwawo mukuyedza kushanda.Kuratidzwa kwenguva refu kunokuvadza hutano hwako, kunyanya kubata kweganda zvakananga kunogona kukonzera kupisa kwakanyanya kwemakemikari uye mavanga asingagumi.

Mumamiriro ezvinhu akajairwa, vadyidzani vemurabhoritari vanogona chete kuderedza njodzi nekuzvishongedza zvizere (kupfeka majasi erabhoritari, magurovhosi nemask).

Uye zano rangu kwauri nderekuti: ndapota tsiva Trizol zvakananga kudzivirira kukuvadzwa kubva kunobva.

1. Chekutanga chizvarwa chigadzirwa cheRNA kutorwa
Buda muna 1987 (Trizol reagents)

mukana:
More kudhirowa kwakakwana

Zvakaipa:
Toxic reagents senge guanidine isothiocyanate-phenol-chloroform inodiwa;
Kushanda zvakaoma;
Inoda kushandisa DNase kubvisa kusvibiswa kweDNA

2.Chizvarwa chechipiri chigadzirwa cheRNA kutorwa
Buda muna 1994 (QIAGEN RNeasy uye yakawanda zvigadzirwa zvepamba)

mukana:
Column centrifugal nzira inogamuchirwa, iyo iri nyore kushanda;

Zvakaipa:
Inoda centrifuge kubvisa tsvina yematishu
Inoda kushandisa DNase kubvisa kusvibiswa kweDNA

3.Chizvarwa chechitatu chigadzirwa cheRNA kutorwa
Kubuda muna 2005 (QIAGEN RNeasy Plus uye ForegeneRNA yekuchenesa yakatevedzana zvigadzirwa)

mukana:
Shandisa zvakananga DNA kubvisa mutsara kubvisa DNA nemarara ematishu panguva imwe chete, kuchengetedza nguva nekuedza;
Yakachengeteka uye iri nyore, hapana chikonzero chekuwedzera sefa column uye DNase kubvisa nyore kusvibiswa uye kudzivirira kusvibiswa kweDNA.

Ngatitarisei misiyano yekushanda pakati pechigadzirwa chechizvarwa chekutanga (Trizol), chizvarwa chechipiri chigadzirwa (RNeasy) uye chechitatu-chizvarwa chigadzirwa (Forgene) cheRNA kubviswa:

Kushanda mumufananidzo kwakajeka: chigadzirwa chechizvarwa chechitatu chine mabhenefiti akajeka

Mazwi matatu chete ekutsanangura: nyore!Fast!Safety

Iyo PK chati yezviyedzo zvabudanokutichizvarwa chekutanga, chechipiri-chizvarwa uye chechitatu-chizvarwa zvigadzirwa zveRNA kubviswa:

1. Electrophoretogram

Chizvarwa chekutanga (1: Trizol)
Chizvarwa chechipiri (2:QIAGEN RNeasy)
Chizvarwa chechitatu (3: QIAGEN RNeasy kuwedzera, 4: Foregene)

Ndeupi rudzi rwechigadzirwa chakanaka, iwe uchaziva nekuenzanisa
RNA yakatorwa uchishandisa iyo Foregene kit
Kwete kungoisa pfungwa huru chete asiwo kuvimbika kwakanaka

2.qPCR girafu (amplification curve graph)

Chizvarwa chekutanga (Trizol)

Chizvarwa chechipiri (sky bluezvinorevaQIAGEN RNeasy-, tsvukuzvinorevaQIAGEN RNeasy +)

Chizvarwa chechitatu (bhuruu zvinoreva Forenene, girini zvinoreva QIAGEN RNeasy plus)

(Cherechedza: Chizvarwa chechipiri cheQIAGEN RNeasy che qPCR girafu chinoreva kuti kititi hachiwedzeri DNase uye hachibvisi kusvibiswa kweDNA. Nokudaro, CT yakakwirira uye mhedzisiro yacho haina kururama; QIAGEN RNeasy + zvinoreva kuti DNase yakawedzerwa uye kusvibiswa kweDNA kwabviswa, uye chigumisiro chacho chakarurama)

Kunyangwe iri electrophoretogram kana yehuwandu PCR amplification curve graph, mhando yezvigadzirwa zveForegene haisi yakaderera kune zvigadzirwa zvinotengeswa kunze kwenyika.