Kugadzirisa Matambudziko ePCR Analyzer: Mibvunzo Inowanzo bvunzwa uye Mhinduro

Vaongorori vePolymerase chain reaction (PCR) zvishandiso zvakakosha mu molecular biology, zvichibvumira vaongorori kuwedzera DNA yekushandisa kubva pakutsvagisa majini kusvika pakuongorora kwekiriniki. Zvisinei, sechero mudziyo wakaoma, PCR analyzer inogona kusangana nematambudziko anokanganisa mashandiro ayo. Chinyorwa chino chinopindura mimwe mibvunzo inowanzo bvunzwa nezveMuchina wekuongorora wePCRkugadzirisa matambudziko uye kunopa mhinduro dzinoshanda kumatambudziko akajairika.

1. Sei maitiro angu ePCR asiri kuwedzera?

Chimwe chezvinetso zvinowanzoitika kune vashandisi ndechekusakwanisa kwePCR reaction kuwedzera DNA yavanoda. Izvi zvinogona kukonzerwa nezvinhu zvakati wandei:

Dhizaini yeprimer isiriyo: Iva nechokwadi chekuti maprimer ako akanyatsoenderana nehurongwa hwaunoda uye ane tembiricha yakakwana yekunyunguduka (Tm). Shandisa zvishandiso zvesoftware padhizaini yeprimer kudzivirira kubatana kusingatsananguriki.

Template DNA Isina Kukwana: Iva nechokwadi chekuti uri kushandisa template DNA yakakwana. Kusakwana kuchaita kuti amplification isashande zvakanaka kana kuti isashande zvachose.

Zvinodzivisa mumuenzaniso: Zvinosvibisa mumuenzaniso zvinogona kudzivirira PCR reaction. Funga nezvekuchenesa DNA yako kana kushandisa imwe nzira yekuburitsa.

Mhinduro: Tarisa dhizaini yako yekutanga, wedzera huwandu hwetemplate, uye iva nechokwadi chekuti sampuro yako haina zvinodzivisa.

2. Sei chigadzirwa changu chePCR chisina saizi isiriyo?

Kana saizi yechigadzirwa chako chePCR isiri sezvaitarisirwa, zvinogona kuratidza dambudziko nemamiriro ezvinhu ekuita kana zvinhu zvakashandiswa.

Kuwedzerwa kwesimba kusingatarisirwi: Izvi zvinogona kuitika kana primer ikabatana nenzvimbo isingatarisirwi. Tarisa kuti ma primer acho akanyatsojeka here uchishandisa chishandiso chakadai seBLAST.

Kupisa Kusina Kunaka: Kana tembiricha yekubatidza yakadzikira zvakanyanya, kubatana kusina kujeka kunogona kuitika. Kugadziriswa kwetembiricha yekubatidza nePCR.

Mhinduro: Simbisa kuti primer chaiyo uye gadzirisa tembiricha ye annealing kuti uvandudze kururama kwezvigadzirwa zvePCR.

3. PCR analyzer yangu inoratidza meseji yekukanganisa. Ndoita sei?

Mashoko ekukanganisa ari paPCR analyzer anogona kutyisa, asi anowanzo kupa zviratidzo zvematambudziko angangoitika.

Matambudziko eCalibration: Iva nechokwadi chekuti PCR analyzer yakaongororwa nemazvo. Kuongororwa nguva dzose kwekugadzirisa nekugadzirisa kwakakosha kuti uwane mhedzisiro chaiyo.

Boka reSoftware: Dzimwe nguva, zvikanganiso zvesoftware zvinogona kukonzera matambudziko. Tangazve komputa yako uye tarisa kana paine zvigadziriso zvesoftware.

MHINDURO: Tarisa bhuku rekushandisa kuti uwane kodhi chaiyo yekukanganisa uye tevera matanho anokurudzirwa ekugadzirisa matambudziko. Kugadzirisa nguva dzose kunogona kudzivirira matambudziko akawanda.

4. Sei mhinduro dzangu dzePCR dzisingaenderane?

Kusaenderana kwePCR kunogona kushungurudza nekuda kwezvikonzero zvakasiyana-siyana:

Hunhu hweReagent: Iva nechokwadi chekuti mareagent ese, kusanganisira maenzyme, mabuffers, uye madNTPs, matsva uye emhando yepamusoro. Mareagents apera kana kuti ane utachiona anogona kukonzera kusiyana.

Kugadziriswa kweThermal Cycler: Kusagadzikana kwemamiriro ekupisa kunogona kukanganisa maitiro ePCR. Gara uchitarisa calibration yethermal cycler.

Mhinduro: Shandisa ma reagents emhando yepamusoro uye gadzirisa thermal cycler yako nguva nenguva kuti uve nechokwadi chekuti mhedzisiro yacho inowirirana.

5. Nzira yekuvandudza sei mashandiro ePCR reaction?

Kuvandudza mashandiro ePCR reactions kunogona kutungamira kugohorera kwakanyanya uye mhedzisiro yakavimbika.

Gadzirisa mamiriro ekuita: Edza uchishandisa huwandu hwakasiyana hwema primers, template DNA uye MgCl2. Maitiro ega ega ePCR angada mamiriro akasiyana kuti ashande zvakanaka.

Shandisa ma enzymes ane kuvimbika kwakanyanya: Kana kururama kwakakosha, funga nezvekushandisa polymerase yeDNA ine kuvimbika kwakanyanya kuderedza zvikanganiso panguva yekuwedzera.

Mhinduro: Ita bvunzo yekuvandudza kuti uwane mamiriro akanakisa ekugadzirisa PCR yako.

Muchidimbu

Kugadzirisa matambudzikoMuchina wekuongorora wePCRBasa rinogona kuva rakaoma, asi kunzwisisa matambudziko akajairika uye mhinduro dzawo kunogona kuwedzera zvakanyanya ruzivo rwako rwePCR. Nekugadzirisa matambudziko aya akajairika, vaongorori vanogona kuvandudza mhedzisiro yePCR uye kuona mhedzisiro yakavimbika mukushandiswa kwebiology yemamorekuru. Kugara uchigadzirisa, kusarudza ma reagents nekungwarira, uye kugadzirisa mamiriro ekuita ndizvo zvakakosha pakubudirira kwePCR analysis.