Xirmooyinka Go'doominta DNA-da bakteeriyada Bakteeriyada Genomic DNA Xirmooyinka Sifaynta
Sharaxaada Qalabka:
Si dhakhso leh uga nadiifi DNA-ga tayada sare leh ee bakteeriyada ee marxaladda koritaanka logarithmic.
◮Wax wasakhaysan RNase ma jiro:Tiirka-DNA-Kaliya ee ay bixiso qalabku waxa uu suurtogal ka dhigayaa in RNA laga saaro DNA-da genomic iyada oo aan lagu darin RNase inta lagu jiro tijaabada, iyada oo ka hortagaysa shaybaarka in uu wasakheeyo RNase dibadda ah.
◮Xawaaraha dheereeya:Foregene Protease wuxuu leeyahay dhaqdhaqaaq ka sarreeya borotiinka la midka ah, wuxuuna si dhakhso ah u dheefshiidaa shaybaarada unugyada;Qalliinku waa sahlan yahay, waxaana hawlgalka soo saarista DNA-ga genomic lagu dhamayn karaa 20-80 daqiiqo gudahood.
◮Ku habboon:Centrifugation-ka waxaa lagu sameeyaa heerkulka qolka, mana jirto baahi heerkul hoose oo 4°C ah ama roobka ethanol ee DNA.
◮Badbaadada:Looma baahna soo saarista reagent organic.
◮Tayada sareDNA-da genomic ee la soo saaray ayaa leh jajabyo waaweyn, ma jiro RNA, ma jiro RNase, iyo maadada ion oo aad u hooseeya, taas oo buuxin karta shuruudaha tijaabooyin kala duwan.
◮Nidaamka Micro-elution:Waxay kordhin kartaa diiradda DNA-da genomic, taas oo ku habboon ogaanshaha hoose ama tijaabinta.
Sharaxaada
Qalabkani wuxuu bixiyaa hab degdeg ah oo sahlan oo laga soo saaro DNA-da genomic ee dhaqamada bakteeriyada (Gram-negative iyo Gram-positive) ilo kala duwan;waxay ka baaraandegi kartaa wax ka yar 3ml ee dhaqamada bakteeriyada ee wajiga koritaanka logarithmic (1×109 bakteeriyada).Qalabka waxa uu la jaanqaadayaa waxtarka sare ee Foregene Protease, kaas oo u oggolaanaya 15-50μg oo DNA-ga tayada sare leh in laga soo saaro 1 saac gudahood.Intaa waxaa dheer, qalabku wuxuu sidoo kale soo saari karaa walxo hidde ah oo aan ahayn genome, sida plasmids, Cosmid, BAC, iwm.
Qalabka DNA-ka kaliya ee silica gel matrix ee loo isticmaalo tiirarka lafdhabarta ayaa ah shay cusub oo shirkadda ah, kaas oo si hufan oo gaar ah u xajin kara DNA-da.Awoodda adsorption ee ugu badan ee DNA waa 80μg.Nidaamka kaydinta gaarka ah iyo nidaamka sare-u-qaadista ayaa kor u qaadi kara ka saarista RNA, borotiinnada wasakhaysan, ions iyo xeryahooda kale ee organic ee unugyada.Qaybta DNA-da ee la soo saaray waa mid weyn, nadiif sare, tayo deggan oo la isku halayn karo, cabbirka jajabka DNA-ga waa mid deggan qiyaastii 23kb.
Tilmaamaha
50 Diyaarin, 100 Diyaarin, 250 Diyaarin
Qaybaha kitka
| Buffer ML1 |
| Buffer ML2 |
| Buffer PW |
| Buffer WB |
| Buffer EB |
| Kahortagga Protease |
| Lysozyme |
| Buffer TE |
| Tiirka-Kaliya DNA |
| Tilmaamaha |
Tilmaamaha & faa'iidooyinka
-Ma jirto wasakheynta RNase: Looma baahna in lagu daro RNase si looga saaro RNA ee DNA-da genomic, iyada oo laga fogaanayo shaybaarka in uu wasakheeyo RNase dibadda ah.
Xawaaraha degdega ah: Foregene Protease waxa uu leeyahay hawl ka sarraysa borotiijiyeyaasha la midka ah oo waxa ay ku dheefi kartaa shay-bakteeriyada jabtay muddo gaaban gudaheed.
-Ku habboonaanta: centrifugation-ka waxaa lagu sameeyaa heerkulka qolka, mana jirto baahi loo qabo 4°C heerkulka hoose ee centrifugation ama roobka ethanol ee DNA.
-Badbaadada: soo saarista reagents organic looma baahna.
-Tayada sare: Qaybaha DNA-da ee la soo saaray waa waaweyn yihiin, ma jiraan RNA, ma jiro RNase, iyo maadada ion aad u hooseeya, taas oo buuxin karta shuruudaha tijaabooyin kala duwan.
Codsiga qalabka
Qalabkani waxa uu ku habboon yahay in lagu sifeeyo DNA-da genomic ee muunadaha soo socda: Bakteeriyada Gram-negative iyo bakteeriyada Gram-positive ee marxaladda koritaanka logarithmic.
Socodka shaqada
Kaydinta iyo nolosha shelf
-Qalabkan waxa lagu kaydin karaa 12 bilood xaaladaha qalalan ee heerkulka qolka (15-25°C);Haddii loo baahdo in lagu kaydiyo wakhti dheer, waxa lagu kaydin karaa 2-8 ° C.
Fiiro gaar ah: Haddii lagu kaydiyo heerkul hooseeya, xalku wuxuu u nugul yahay roobab.Kahor intaan la isticmaalin, hubso inaad geliso xalka qalabka heerkulka qolka muddo wakhti ah.Haddii loo baahdo, ku sii kululee biyaha qubeyska 37°C 10 daqiiqo si aad u milanto daadka, oo ku qas kahor isticmaalka.
-Xalka Foregene Protease wuxuu leeyahay qaacido gaar ah, kaas oo firfircoon marka lagu kaydiyo heerkulka qolka muddo dheer (3 bilood);marka lagu kaydiyo 4°C, dhaq-dhaqaaqiisa iyo degenaanshihiisa ayaa fiicnaan doona, sidaa awgeed waxa lagu talinayaa in lagu kaydiyo 4°C, xasuuso inaadan ku haynin -20°C.
-Lysozyme waa budo qalalan, oo firfircoon marka lagu kaydiyo 2-8 ° C muddo dheer (3 bilood).Kaydinta dheer, fadlan ku kaydi heerkulka -20°C.
