Binciken da ke gaba na yiwuwar matsalolin a cikinBukalswab/FTA card DNA extraction is helpful to your experiment. In addition, for other experimental or technical problems in addition to operating instructions and problem analysis, we have dedicated technical support to help you. If you have any needs, please contact us at: 028-83360257 or E-mali:Tech@foregene.com.

Rukunin tsarkakewa ya toshe

A cikin wannan kit ɗin, a cikin aikin hakar DNA na genomic, ginshiƙin tsarkakewa yana tallata kai tsaye a kan cakudawar enzymatic lysis na samfurin ba tare da matakin centrifugation ba, kuma ana iya toshe ginshiƙin tsarkakewa saboda rashin cikar enzymization da babban danko na samfurin.

Dalilan da ke yiwuwa su ne kamar haka:

1. Rashin cikakkiyar narkewar enzymatic na samfuran nama.

Shawarwari: Za'a iya tsawaita lokacin sarrafa samfurin na Foregene Protease daidai ko kuma ana iya ɗaukar maɗaukaki bayan centrifugation a 12,000 rpm (~ 13,400)× g) da 5 min.

2. Yin amfani da samfurori masu yawa ko manyan kyallen takarda.

Shawarwari: Yana da kyau kada ku wuce 1Bukal swab a cikin samfurin;idan samfurin ya yi girma, ƙara yawan adadin Buffer ST1, Foregene Protease, buffer ST2 daidai da haka.

3. Samfurin danko ya yi yawa.

Shawarwari: Za'a iya narkar da samfurori daidai da 10mM na Tris-HCl kafin cirewar DNA na kwayoyin halitta.

4. An tsotse guntuwar katin jini.

Shawarwari: Lokaci na wucin gadi na mataki na 6 na cirewar jini (Katin FTA) na iya tsawaita yadda ya kamata.

Ƙananan yawan amfanin ƙasa ko babu DNA

Sau da yawa akwai nau'o'i iri-iri waɗanda ke shafar yawan amfanin DNA na genomic, ciki har da asalin samfurin, yanayin ajiyar samfurin, shirye-shiryen samfurin, magudi, da dai sauransu.

Ba za a iya samun DNA na genomic yayin hakar ba

Abubuwan da za su iya haifar da su sune kamar haka:

1. Rashin kyaun adana samfurori ko ajiya na dogon lokaci yana haifar da lalata DNA na kwayoyin halitta.

Shawarwari: Ya kamata a yi amfani da swabs na baka da sabon samfuri, kuma ba shi da kyau a yi amfani da swabs da aka adana don ayyukan hakar DNA;samfuran tabo na jini yakamata su tabbatar da cewa ingancin ya cancanta kuma lokacin ajiya bai kamata ya yi tsayi da yawa ba.

2. Yin amfani da nama kaɗan kaɗan na iya haifar da rashin hako DNA ɗin da ya dace.

Shawarwari: Bibukkal umarnin samfurin swab a cikin jagorar aiki, kuma a goge sau da yawa sosai yadda za a iya haɗa isassun sel zuwa swab na baka don hakar DNA na genomic;don hakar samfurin tabo na jini, ana iya ƙara yankin yankan jini yadda ya kamata.

3. Foregene Protease ana kiyaye shi ba daidai ba, yana haifar da raguwa a cikin ayyukansa ko rashin kunnawa.

Shawarwari: Tabbatar da yanayin ajiya nada Foregene Protease ko maye gurbin shi da sabon Foregene Protease don amsawar enzymatic.

4. Adana rashin dacewa na kit ko lokacin ajiya yayi tsayi da yawa, yana haifar da gazawar wasu abubuwan da ke cikin kit ɗin.

Shawarwari: Sayi sabuwaBukal swab DNAkaɗaici kit don hanyoyin da suka danganci.

5. Buffer WB baya ƙara cikakken ethanol.

Shawarwari: Tabbatar da cewa buffer WB yana ƙara madaidaicin ƙarar cikakken ethanol.

6. A eluent ba daidai ba a kara zuwa fim din silicone.

Shawarwari: Ƙara 65°Cpre-warmed eluent ya sauko zuwa tsakiyar silicone membrane kuma ya bar a dakin da zafin jiki na 5 min don ƙara haɓaka haɓakar haɓakawa.

LDNA na kwayoyin halitta ware

Dalilan da ke yiwuwa su ne kamar haka:

1. Rashin kyaun adana samfurori ko ajiya na dogon lokaci yana haifar da lalata DNA na kwayoyin halitta.

Shawarwari: An fi dacewa da swabs na baki an gwada sabon samfuri, kuma ba za a yi amfani da swabs ɗin da aka adana ba don hakar DNA na kwayoyin halitta.

2. Idan adadin samfurin nama ya yi ƙanƙanta, abin da DNA ɗin da aka fitar zai ragu.

Shawarwari: Bi umarnin samfurin swab na baka a cikin jagorar aiki, shafa sau da yawa sosai yadda za a iya haɗa isassun sel zuwa swab na baka don hakar DNA na kwayoyin halitta.

3. Foregene Protease ana kiyaye shi ba daidai ba, yana haifar da raguwa a cikin ayyukansa ko rashin kunnawa.

Shawarwari: Tabbatar da yanayin ajiya nathe Foregene Protease ko maye gurbin shi da sabo Foregene Protease don amsawar enzymatic.

4. Matsaloli masu ma'ana.

Shawarwari: Yi amfani da Buffer EB don haɓakawa;idan kuna amfani da ddH₂O ko wasu ɓarna, tabbatar da cewa pH na eluate yana tsakanin 7.0-8.5.

5. The eluate ba a daidai ƙara dropwise.

Shawarwari: Ƙara digo na eluent zuwa tsakiyar membrane na silicone kuma barin a dakin da zafin jiki na 5 min don ƙara haɓaka haɓakawa.

6. Ruwan ruwa yana taruwa kadan.

Shawarwari: Yi amfani da eluent don haɓakar DNA na genomic kamar yadda ake buƙata a cikin umarnin, aƙallaba kasa ba fiye da 15μl.

Loya tsarki of kwayoyin halittar DNAware

Ƙananan tsaftar DNA na genomic na iya haifar da gazawa ko sakamako mara gamsarwa na gwaje-gwaje na ƙasa, kamar: enzymes ba za a iya buɗewa ba, PCR ba zai iya samun gutsuttsarin jinsin sha'awa ba, da sauransu.

Abubuwan da za su iya haifar da su sune kamar haka:

1. Rashin gurɓatawar sinadarin heteroprotein, gurɓacewar RNA.

Bincike: Ba a wanke ginshiƙin tsarkakewa ta amfani da Buffer PW;Ba a wanke ginshiƙin tsarkakewa na Buffer PW ba ta amfani da madaidaicin saurin centrifugal.

Shawarwari: Tabbatar cewa babu hazo a cikin supernatant kafin ƙara ethanol;tabbatar da wanke ginshiƙin tsarkakewa bisa ga umarnin, kuma ba za a iya barin wannan matakin ba.

2. Rashin tsarkin ion.

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

Shawarwari: Tabbatar wanke Buffer WB sau 2 kamar yadda aka umarce shi don cire ragowar ions gwargwadon yiwuwa.

3. RNA enzyme gurbatawa.

Nazari: An ƙara RNases na ƙasashen waje zuwa buffer;Aikin wanke kayan buffer PW bai yi daidai ba, yana haifar da ragowar RNase, yana shafar ayyukan gwajin RNA na ƙasa, kamar rubutun in vitro.

Shawarwari: Foregene jerin nucleic acidisolation kits na iya cire RNA ba tare da ƙarin ƙarin RNase ba,haka Buccal Swab/FTA Katin DNA Keɓewa Kitbukata't ƙara RNase;Tabbatar ku bi umarnin Buffer PW ginshiƙin tsarkakewa, kuma ba za a iya barin wannan matakin ba.

4. Ethanol ragowar.

Nazari: Buffer WB bai yi fankon bututu ba bayan wanke ginshiƙin tsarkakewa.

Shawarwari: Yi daidaitaccen aikin santsi na bututu bisa ga umarnin.

5. Sauran kazanta.

Bincike: Samfuran da aka ajiye ko samfurori na musamman ba a riga an tsara su ba.

Shawarwari: Gyara samfurin sosai kamar yadda aka umarce shi.