Buccal Swab/FTA Katin DNA Keɓe Kit ɗin Cirar DNA na Genomic DNA ko Kayan Tsarkakewa daga Buccal Swabs
Bayanin Kit:
Da sauri tsarkake high quality-genomic DNA daga buccal swab/FTA Katin samfuran.
◮Babu gurɓatar RNase:Rukunin DNA-Only da kit ɗin ya samar yana ba da damar cire RNA daga DNA ɗin kwayoyin halitta ba tare da ƙara RNase ba yayin gwajin, yana hana dakin gwaje-gwaje daga kamuwa da RNase na waje.
◮Gudun sauri:Foregene Protease yana da ayyuka mafi girma fiye da irin wannan proteases, kuma yana narkar da samfuran nama da sauri;aikin yana da sauƙi, kuma ana iya kammala aikin hakar DNA na genomic a cikin mintuna 20-80.
◮Dace:Ana yin centrifugation a dakin da zafin jiki, kuma babu buƙatar ƙananan zafin jiki na 4 ° C ko hazo na ethanol na DNA.
◮Tsaro:Ba a buƙatar hakar reagents na halitta.
◮Babban inganci:DNA ɗin da aka fitar yana da manyan gutsuttsura, babu RNA, babu RNase, da ƙarancin ion abun ciki, wanda zai iya biyan buƙatun gwaje-gwaje daban-daban.
◮Tsarin Micro-elution:Yana iya ƙara ƙaddamar da DNA na genomic, wanda ya dace don ganowa ko gwaji.
Bayani
Wannan kit ɗin yana ba da ingantacciyar hanya da sauri don samun babban tattarawar kwayoyin halittar DNA daga buccal swabs da Katin FTA (tabobin jini).Amfani da kamfaninmu's musamman DNA-kawai silica membrane spin column da dabara, hade tare da Foregene Protease, babban maida hankali, ingantaccen DNA na genomic ana iya fitar dashi cikin mintuna 80.Ƙaƙwalwar ƙaramin ginshiƙin tsarkakewa da aka ƙera na musamman yana ɗaure DNA na genomic, kuma DNA ɗin za a iya ɓarna da ƙaramin adadi (15μl) tsarin elution don ƙara yawan tattarawar DNA ɗin da aka samu, wanda ya dace don ganowa ko gwaji.Kit ɗin na iya aiwatar da samfurori ɗaya ko fiye a lokaci ɗaya, kuma tsarin tsarkakewa baya buƙatar hakar abubuwan halitta kamar phenol, chloroform, da isopropanol mai cin lokaci ko hazo na ethanol, kuma aikin yana da sauƙi kuma yana adana lokaci.
Ƙayyadaddun bayanai
50 Shirye-shirye
Abubuwan Kit
| Farashin ST1 |
| Farashin ST2 |
| Acrylamide Linear |
| Farashin PW |
| Buffer WB |
| Buffer EB |
| Maganin Protease |
| Rukunin DNA-Kawai |
| Umarni |
Fasaloli & fa'idodi
-Babu gurɓatawar RNase: Rukunin DNA-kawai wanda kit ɗin ya samar yana ba da damar cire RNA daga DNA na kwayoyin halitta ba tare da ƙara RNase ba yayin gwajin, yana guje wa gurɓatar da dakin gwaje-gwaje ta RNase na waje.
-Saurin sauri: Foregene Protease yana da ayyuka mafi girma fiye da nau'ikan proteases iri ɗaya, narkar da samfuran da sauri;aiki mai sauƙi.
-Mafi dacewa: Ana yin centrifugation a dakin da zafin jiki, kuma babu buƙatar 4 ° C ƙananan zafin jiki ko hazo na ethanol na DNA.
-Safety: Ba a buƙatar hakar reagent na halitta.
-Maɗaukakiyar inganci: DNA ɗin da aka fitar yana da manyan gutsuttsura, babu RNA, babu RNase, da ƙarancin ion abun ciki, wanda zai iya biyan buƙatun gwaje-gwaje daban-daban.
-Micro-elution tsarin: Yana iya ƙara maida hankali na genomic DNA, wanda ya dace don gano ƙasa ko gwaji.
Kit aikace-aikace
Ya dace da tsarkakewa na DNA na genomic daga samfurori masu zuwa: buccal swabs, FTA Card (tabon jini).
Adana da Rayuwar Shelf
-Wannan kit ɗin za a iya adana shi na tsawon watanni 12 a ƙarƙashin busassun yanayi a cikin zafin jiki (15-25 ° C);Idan ana buƙatar adana na dogon lokaci, ana iya adana shi a zazzabi na 2-8 ° C.
Lura: Idan an adana shi a ƙananan zafin jiki, maganin yana da sauƙi ga hazo.Kafin amfani, tabbatar da sanya maganin a cikin kit a cikin zafin jiki na wani lokaci.Idan ya cancanta, preheta shi a cikin ruwan wanka na 37 ° C na tsawon minti 10 don narkar da hazo, kuma a gauraya shi kafin amfani.
-Foregene Protease bayani yana da tsari na musamman, wanda ke aiki lokacin da aka adana shi a dakin da zafin jiki na dogon lokaci (watanni 3);Ayyukansa da kwanciyar hankali za su fi kyau idan an adana su a 4 ° C, don haka ana bada shawara don adana shi a 4 ° C, ku tuna kada ku ajiye shi a -20 ° C.
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 supernatant bayan centrifugation a 12,000 rpm (~ 13,400 × g) na 5 min.
2. Yin amfani da samfurori masu yawa ko manyan kyallen takarda.
Shawarwari: Zai fi kyau kada ku wuce 1 Buccal 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: Bi umarnin samfurin swab na buccal a cikin jagorar aiki, kuma a goge sau da yawa sosai yadda za a iya haɗa isassun sel zuwa swab na baka don cirewar DNA;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 na Foregene Protease ko maye gurbin shi da sabon Protease na Foregene 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 sabon Buccal swab DNA keɓancewar kayan don hanyoyin da ke da alaƙa.
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 °C pre-warmed eluent ya sauke zuwa tsakiyar membrane na silicone kuma barin a dakin da zafin jiki na 5 min don ƙara haɓaka haɓakawa.
Ƙarƙashin amfanin gona na DNA keɓe
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 na Foregene Protease ko maye gurbin shi da sabon Protease na Foregene 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, aƙalla ƙasa da μl 15.
Low tsarki na genomic DNA ware
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 keɓancewar acid nucleic acid na iya cire RNA ba tare da ƙarin ƙarin RNase ba, don haka buccal Swab/FTA Katin DNA keɓe Kit ɗin baya buƙatar 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.
