Jagorar Binciken Matsala

The following is an analysis of the problems that may be encountered in the extraction of plant genomic DNA, hoping to be helpful to your experiments. In addition, for other experimental or technical problems other than operation instructions and problem analysis, we have dedicated technical support to help you. If you have any needs, please contact us: 028-83360257 or E-mali: Tech@foregene.com.

Ƙananan yawan amfanin ƙasa ko babu DNA

Yawancin abubuwa da yawa waɗanda ke shafar yawan amfanin DNA na genomic, gami da tushen samfurin, shekarun samfurin, yanayin ajiyar samfurin, da aiki.

Ba a iya samun DNA na genomic yayin hakar

1. Ana adana samfuran nama ba daidai ba ko adana su na dogon lokaci, yana haifar da lalacewar DNA na genomic.

Shawarwari: Ajiye samfuran nama a cikin ruwa nitrogen ko -20°C;gwada amfani da sabbin samfuran da aka tattara don hakar DNA na kwayoyin halitta.

2. Ƙananan adadin adadin samfurin zai iya sa ba za a fitar da DNA ɗin da ya dace ba.

Shawarwari: Don samfuran nama waɗanda aka adana na dogon lokaci ko suna da mummunan lalata DNA, adadin nama za a iya ƙara yadda ya kamata don fitar da DNA mai yawa.Ana iya ƙayyade adadin samfurin bisa ga bukatun DNA, amma samfurin sabo bai kamata ya wuce 100mg ba, kuma busassun samfurin kada ya wuce 30mg.

3. Samfurin ba a ƙasa tare da nitrogen na ruwa ba ko sanya shi na dogon lokaci bayan nitrogen na ruwa.

Shawara: Yayin hakar DNA, samfurin yana buƙatar zama cikakke tare da nitrogen na ruwa don karya bangon tantanin halitta;bayan niƙa, don Allah canja wurin samfurin foda zuwa PL1 preheated a 65°C da wuri-wuri (da zarar foda na ƙasa ya narke, DNA na genomic zai fara raguwa da sauri).

4. Rashin ingantaccen ajiya na Foregene Protease yana haifar da raguwa ko aiki mara aiki.

Shawarwari: Tabbatar da yanayin ajiya na Foregene Protease ko musanya shi da sabon Foregene Protease don enzymatic hydrolysis.

5. Ana adana kayan da ba daidai ba ko adana su na dogon lokaci, yana haifar da gazawar wasu abubuwan da ke cikin kit ɗin.

Shawarwari: Sayi sabon kayan hako kwayoyin halittar DNA na shuka don ayyuka masu alaƙa.

6. Yin amfani da kayan da bai dace ba.

Shawarwari: Sayi Kit ɗin keɓewar DNA na Tsire-tsire da aka keɓe don samfurori don hakar da tsarkakewar DNA na kwayoyin halitta.

7. Buffer WB ba tare da ƙara am ethanol.

Shawarwari: Tabbatar da ƙara madaidaicin ƙarar cikakken ethanol zuwa Buffer WB.

8. Ba a ɗigo da eluent a jikin siliki ba daidai ba.

Shawarwari: Ƙara zafin zafin jiki a 65℃dropwise zuwa tsakiyar silica gel membrane, da kuma bar shi a dakin zafin jiki na 5 minutes don ƙara da elution yadda ya dace.

Cire don samun ƙananan amfanin gona na DNA

1. Ana adana samfurin da ba daidai ba ko adana shi na dogon lokaci, yana haifar da lalacewar DNA na kwayoyin halitta.

Shawarwari: Ajiye samfuran nama a -20℃;gwada amfani da sabbin samfuran nama da aka tattara don hakar DNA na kwayoyin halitta.

2. Idan adadin samfuran nama ya yi ƙanƙanta, DNA ɗin da aka fitar zai zama ƙasa.

Shawara: Wasu samfuran tsire-tsire suna da wadataccen ruwa, kamar tsire-tsire na ruwa kamar algae, da dai sauransu, ana iya ƙara yawan adadin da ya dace ko kuma ruwan zai iya bushewa kaɗan kafin aikin.

3. Samfurori ba su da ƙasa sosai tare da nitrogen na ruwa ko an bar su a dakin da zafin jiki na dogon lokaci bayan niƙa.

Shawara: Dole ne niƙa nitrogen ta ruwa ya isa, kuma bangon samfurin ya kamata a karye gwargwadon yiwuwa;nan da nan bayan niƙa, samfurin foda ya kamata a canza shi zuwa 65℃preheated Buffer PL1 don mataki na gaba.

4. Rashin amfani da kayan aiki daidai.

Shawarwari: Yi amfani da keɓaɓɓen Kit ɗin keɓewar DNA don cirewa da tsarkake halittar DNA.

5. Rashin ingantaccen ajiya na Foregene Protease yana haifar da raguwa ko aiki mara aiki.

Shawarwari: Tabbatar da yanayin ajiya na Foregene Protease ko musanya shi da sabon Foregene Protease don enzymatic hydrolysis.

6. Matsala mai haske

Shawarwari: Da fatan za a yi amfani da Buffer EB don haɓakawa;idan kuna amfani da ddH₂O ko wasu ɓarna, tabbatar da cewa pH na eluent yana tsakanin 7.0-8.5.

7. Ba a ɗigowar ƙuruciya daidai

Shawarwari: Da fatan za a ƙara digo na elution zuwa tsakiyar silica membrane kuma bar shi a cikin zafin jiki na tsawon mintuna 5 don ƙara haɓaka haɓaka.

8. Ƙarfin eluent ya yi ƙanƙanta sosai

Shawara: Da fatan za a yi amfani da eluent don haɓakar DNA na genomic bisa ga umarnin, aƙalla ƙasa da 100μl.

Cire genomic DNA tare da ƙarancin tsabta

Ƙananan tsarki na DNA na genomic zai haifar da gazawa ko mummunan sakamako na gwaje-gwaje na ƙasa, kamar: ba za a iya yanke enzyme ba, kuma PCR ba zai iya samun gutsuttsarin kwayoyin halitta ba.

1. Gurɓatar furotin daban-daban, gurɓataccen RNA.

Bincike: Ba a yi amfani da Buffer PW don wanke ginshiƙi ba;Ba a yi amfani da Buffer PW ba don wanke ginshiƙi a daidai saurin ɗawainiya.

Shawara: gwada ƙoƙarin tabbatar da cewa babu hazo a cikin maɗaukaki lokacin da aka wuce ta cikin ginshiƙi;tabbatar da wanke ginshiƙin tsarkakewa tare da Buffer PW bisa ga umarnin, kuma wannan matakin ba za a iya tsallake shi ba.

2. Rashin tsarkin ion.

Nazari: An cire ginshiƙin wankin Buffer WB ko kuma an wanke shi sau ɗaya kawai, yana haifar da gurɓataccen gurɓataccen ionic.

Shawarwari: Tabbatar yin wanka sau biyu tare da Buffer WB bisa ga umarnin don cire ragowar ions gwargwadon yiwuwa.

3. RNase gurbatawa.

Analysis: Exogenous RNase yana ƙara zuwa majigi;Aikin wankin da ba daidai ba a cikin Buffer PW zai haifar da ragowar RNase kuma ya shafi ayyukan gwajin RNA na ƙasa, kamar rubutun in vitro.

Shawara: Foregene jerin abubuwan hakar acid nucleic acid na iya cire RNA ba tare da ƙarin RNase ba, kuma duk reagents a cikin Kit ɗin keɓewar DNA ba sa buƙatar RNase;tabbatar da wanke ginshiƙin tsarkakewa tare da Buffer PW bisa ga umarnin, kuma wannan matakin ba za a iya tsallake shi ba.

4. Ragowar Ethanol.

Nazari: Bayan wanke ginshiƙin tsarkakewa tare da Buffer WB, ba a yi wani fanni na bututu ba.

Shawarwari: Bi umarnin don daidaitaccen bututu centrifugation.

Jagoran Jagora:

Manual Umarnin Keɓewar Tsibirin DNA