Buccal Swab/FTA Kaadị DNA kewapụ Kit Genomic DNA mmịpụta ma ọ bụ ihe dị ọcha sitere na Buccal Swabs
Nkọwa ngwa:
Mee ngwa ngwa DNA genomic dị elu site na nlele kaadị swab/FTA.
◮Enweghị mmetọ RNase:Ogidi DNA-naanị nke ngwa ahụ na-enye na-eme ka o kwe omume iwepụ RNA na DNA genomic na-agbakwunyeghị RNase n'oge nnwale ahụ, na-egbochi ụlọ nyocha ahụ site na RNase exogenous metọọ.
◮Ọsọ ọsọ:Foregene Protease nwere ọrụ dị elu karịa proteases yiri ya, ma na-agbari ihe nlele anụ ahụ ngwa ngwa;arụ ọrụ ahụ dị mfe, yana ọrụ mmịpụta DNA nke genomic nwere ike mechaa n'ime nkeji 20-80.
◮Dị mma:A na-eme centrifugation na okpomọkụ nke ụlọ, ọ dịghịkwa mkpa maka 4 Celsius C obere okpomọkụ centrifugation ma ọ bụ ethanol precipitation nke DNA.
◮Nchekwa:Ọ dịghị organic reagent mmịpụta a chọrọ.
◮Ịdị mma Dị elu:DNA genomic nke ewepụtara nwere nnukwu iberibe, enweghị RNA, enweghị RNase, yana ọdịnaya ion dị oke ala, nke nwere ike izute ihe nyocha dị iche iche chọrọ.
◮Sistemụ Micro-elution:Ọ nwere ike ịbawanye ntinye nke DNA genomic, nke dị mma maka nchọpụta ala ma ọ bụ nnwale.
Nkọwa
Ngwa a na-enye usoro dị mma na ngwa ngwa iji nweta DNA genomic dị elu site na swabs buccal na kaadị FTA ( ntụpọ ọbara).Iji ụlọ ọrụ anyị's pụrụ iche DNA-naanị silica akpụkpọ anụ spin kọlụm na usoro, jikọtara ya na Foregene Protease, elu-ntụkwasị obi, elu-edu genomic DNA nwere ike isi na 80 nkeji.Obere kọlụm a na-eme ka ọ dị ọcha ahaziri ahazi na-ejikọ DNA genomic, na DNA nwere ike were obere ego wepụta ya.15μl) usoro elution iji mee ka ntinye nke DNA genomic enwetara, nke dị mma maka nchọpụta ala ma ọ bụ nnwale.Ngwa ahụ nwere ike ịhazi otu ma ọ bụ karịa sample n'otu oge, na usoro nchacha anaghị achọ mmịpụta nke ihe ndị dị ndụ dị ka phenol, chloroform, na isopropanol ma ọ bụ ethanol na-ewe oge, na ọrụ ahụ dị mfe ma na-azọpụta oge.
Nkọwapụta
50 nkwadebe
Ngwa ngwa
| Ihe nchekwa ST1 |
| Ihe nchekwa ST2 |
| Acrylamide linear |
| Ihe nchekwa PW |
| Ihe nchekwa WB |
| Ihe nchekwa EB |
| Protease nke mbụ |
| DNA-naanị kọlụm |
| Ntuziaka |
Atụmatụ & uru
-Ọnweghị mmetọ RNase: Otu kọlụm DNA-naanị nke ngwa ahụ na-enye na-eme ka o kwe omume iwepu RNA na DNA genomic na-agbakwunyeghị RNase n'oge nnwale ahụ, na-ezere ụlọ nyocha ahụ ka ọ ghara imerụ ya site na RNase dịpụrụ adịpụ.
-Ọsọ ọsọ ọsọ: Foregene Protease nwere ọrụ dị elu karịa proteases yiri ya, na-agbari sample ngwa ngwa;ọrụ dị mfe.
-Ọ dị mma: A na-eme centrifugation na ụlọ okpomọkụ, ọ dịghịkwa mkpa maka 4 Celsius C dị ala ma ọ bụ ethanol precipitation nke DNA.
-Safety: Ọ dịghị organic reagent mmịpụta a chọrọ.
-Ọdịmma dị elu: DNA genomic amịpụtara nwere nnukwu iberibe, enweghị RNA, enweghị RNase, yana ọdịnaya ion dị oke ala, nke nwere ike izute ihe achọrọ nnwale dị iche iche.
Sistemụ Micro-elution: Ọ nwere ike ịbawanye ntinye nke DNA genomic, nke dị mma maka nchọpụta ala ma ọ bụ nnwale.
Ngwa ngwa
Ọ dabara maka ịdị ọcha nke DNA genomic site na ihe atụ ndị a: swabs buccal, Kaadị FTA (ncha ọbara).
Nchekwa na Shelf ndụ
-Enwere ike ịchekwa ngwa a maka ọnwa 12 n'okpuru ọnọdụ akọrọ na ụlọ okpomọkụ (15-25 ° C);Ọ bụrụ na achọrọ ịchekwa ya ogologo oge, enwere ike ịchekwa ya na 2-8 Celsius.
Rịba ama: Ọ bụrụ na echekwara ya na obere okpomọkụ, ngwọta na-adịkarị mfe mmiri ozuzo.Tupu ojiji, jide n'aka na ị ga-etinye ihe ngwọta na kit na ụlọ okpomọkụ maka oge.Ọ bụrụ na ọ dị mkpa, kpoo ya na mmiri ịsa mmiri 37 Celsius C maka nkeji 10 iji gbazee mkpọda ahụ, ma gwakọta ya tupu ejiri ya.
-Foregene Protease ngwọta nwere usoro pụrụ iche, nke na-arụ ọrụ mgbe echekwara na ụlọ okpomọkụ ruo ogologo oge (ọnwa 3);ọrụ ya na nkwụsi ike ya ga-aka mma ma ọ bụrụ na echekwara ya na 4 Celsius C, ya mere, a na-atụ aro ka ịchekwa ya na 4 Celsius C, cheta ka ọ ghara idobe ya na -20 Celsius.
Akpọchiela kọlụm nchacha
N'ime ngwa a, na ọrụ mmịpụta DNA nke genomic, a na-etinye kọlụm dị ọcha ozugbo na ngwakọta enzymatic lysis sample na-enweghị nzọụkwụ centrifugation, na kọlụm dị ọcha nwere ike igbochi n'ihi enzymization ezughị ezu na viscosity dị elu nke ihe nlele ahụ.
Ihe ndị a nwere ike ịkpata bụ ndị a:
1. Enzymatic mgbaze nke anụ ahụ ezughị ezu.
Nkwanye: Enwere ike ịgbatị oge nhazi ihe nlele nke Foregene Protease nke ọma ma ọ bụ enwere ike iwere ihe karịrị centrifugation na 12,000 rpm (~ 13,400 × g) maka 5 min.
2. Iji ihe nlele anụ ahụ ma ọ bụ nnukwu anụ ahụ mee ihe nke ukwuu.
Nkwanye: Ọ kacha mma ka ị ghara ịgafe 1 Buccal swab na nlele;Ọ bụrụ na ihe nlele ahụ buru ibu, mụbaa dose nke Buffer ST1, Foregene Protease, buffer ST2 otu a.
3. Ihe nlele viscosity dị oke elu.
Nkwanye: Enwere ike were 10mM nke Tris-HCl gbarie ihe nlele nke ọma tupu mmịpụta DNA genomic.
4. Arachala iberibe kaadị ọbara.
Nkwanye: Enwere ike ịgbatị oge centrifugation na-agafe agafe nke nzọụkwụ 6 nke ntụpọ ọbara (FTA Card) mmịpụta mkpụrụ ndụ ihe nketa nke ọma.
Mkpụrụ dị ala ma ọ bụ enweghị DNA
A na-enwekarị ihe dị iche iche nke na-emetụta mkpụrụ DNA genomic, gụnyere ihe atụ sitere, ọnọdụ nchekwa ihe nlele, nkwadebe nlele, nhazi, wdg.
Enweghị ike ịnweta DNA genomic n'oge mmịpụta
Ihe nwere ike ịkpata ya bụ ndị a:
1. Ichekwa ihe nlele ma ọ bụ nchekwa na-ezighi ezi maka ogologo oge na-eduga na mmebi DNA genomic.
Nkwanye: Ọkacha mma swabs ọnụ ka a ga-enyocha ya nke ọma, ọ bụghịkwa mma iji swabs echekwara maka ọrụ mmịpụta DNA genomic;Ihe nlele ntụpọ ọbara kwesịrị ijide n'aka na ogo ahụ tozuru oke yana oge nchekwa ekwesịghị ịdị ogologo.
2. Ojiji nke anụ ahụ dị nta nwere ike ime ka enweghị mmịpụta nke DNA genomic kwekọrọ.
Nkwanye: Soro ntuziaka sampling buccal swab na ntuziaka ọrụ, wee hichaa ugboro ole o kwere mee ka enwere ike itinye mkpụrụ ndụ zuru oke na swab ọnụ maka mmịpụta DNA genomic;maka mmịpụta ntụpọ ọbara, enwere ike ịbawanye ebe mbepụ ntụpọ ọbara nke ọma.
3. A na-echekwa Foregene Protease n'ụzọ na-ekwesịghị ekwesị, na-ebute mbelata ọrụ ya ma ọ bụ arụ ọrụ ya.
Nkwanye: Kwado ọnọdụ nchekwa nke Foregene Protease ma ọ bụ dochie ya na Foregene Protease ọhụrụ maka mmeghachi omume enzymatic.
4. Nchekwa ezighi ezi nke ngwa ma ọ bụ oge nchekwa dị ogologo, na-ebute ọdịda nke ụfọdụ ihe dị na ngwa ahụ.
Nkwanye:zụta ihe ọhụrụ Buccal swab DNA maka usoro ndị metụtara ya.
5. Buffer WB anaghị etinye ethanol zuru oke.
Nkwanye: Kwenye na ihe nchekwa WB na-agbakwụnye olu ziri ezi nke ethanol zuru oke.
6. A naghị agbakwunye eluent nke ọma na ihe nkiri silicone.
Nkwanye: Tinye 65 Celsius C pre-warmed eluent na-adaba n'etiti akpụkpọ ahụ silicone wee hapụ n'ime ụlọ okpomọkụ maka 5 min iji bulie arụmọrụ elution.
Genomic DNA dị ala dịpụrụ adịpụ
Ihe ndị a nwere ike ịkpata bụ ndị a:
1. Ichekwa ihe nlele ma ọ bụ nchekwa na-ezighi ezi maka ogologo oge na-eduga na mmebi DNA genomic.
Nkwanye: Ọkachamma swabs ọnụ ka a na-enyocha nke ọhụrụ, na swabs echekwara ekwesịghị iji maka mmịpụta DNA genomic.
2. Ọ bụrụ na ọnụọgụ anụ ahụ dị ntakịrị, ọdịnaya DNA genomic amịpụtara ga-adị ntakịrị.
Nkwanye: Soro ntuziaka nlele ọnụ swab dị na ntuziaka ịwa ahụ, na-ehichapụ ọtụtụ oge ka enwere ike itinye mkpụrụ ndụ zuru oke na swab ọnụ maka mmịpụta DNA genomic.
3. A na-echekwa Foregene Protease n'ụzọ na-ekwesịghị ekwesị, na-ebute mbelata ọrụ ya ma ọ bụ arụ ọrụ ya.
Nkwanye: Kwado ọnọdụ nchekwa nke Foregene Protease ma ọ bụ dochie ya na Foregene Protease ọhụrụ maka mmeghachi omume enzymatic.
4. nsogbu ndị na-egbuke egbuke.
Nkwanye: Jiri Buffer EB maka elution;Ọ bụrụ na ị na-eji ddH2O ma ọ bụ eluent ndị ọzọ, gosi na pH nke eluate dị n'etiti 7.0-8.5.
5. A naghị agbakwunye eluate nke ọma.
Nkwanye: Tinye eluent tụlee n'etiti akpụkpọ ahụ silicone wee hapụ na ọnụ ụlọ okpomọkụ maka 5 min iji welie elution arụmọrụ.
6. Mmiri elution na-akwakọba ntakịrị.
Nkwanye: Jiri eluent maka elution DNA genomic dị ka achọrọ na ntuziaka, opekata mpe ihe na-erughị 15 μl.
Obere ịdị ọcha nke DNA genomic dịpụrụ adịpụ
Ọdịdị DNA dị ala dị ala nwere ike iduga ọdịda ma ọ bụ nsonaazụ na-adịghị mma nke nyocha ndị dị n'okpuru, dị ka: enweghị ike imeghe enzymes, PCR enweghị ike ịnweta mkpụrụ ndụ mkpụrụ ndụ nke mmasị, wdg.
Ihe nwere ike ịkpata ya bụ ndị a:
1. Mmetọ heteroprotein, mmetọ RNA.
Nyocha: A naghị akwọcha kọlụm dị ọcha site na iji Buffer PW;A saghị kọlụm nchacha nke Buffer PW site na iji ọsọ centrifugal ziri ezi.
Nkwanye: Gbaa mbọ hụ na ọ dịghị mmiri ozuzo na supernatant tupu ịgbakwunye ethanol;jide n'aka na ị ga-asacha kọlụm dị ọcha dịka ntuziaka ahụ si dị, na enweghị ike ịhapụ nzọụkwụ a.
2. Mmetọ ion adịghị ọcha.
Nyocha: Ewepụla ma ọ bụ sachaa kọlụm ịsacha ihe nchekwa nke WB naanị otu ugboro, butere mmetọ ionic fọdụrụnụ.
Nkwanye: Jide n'aka na ị ga-asa Buffer WB ugboro 2 ka enyere ya ka iwepu ion fọdụrụ ka o kwere mee.
3. RNA enzyme mmetọ.
Nyocha: Agbakwunyere RNases nke mba ọzọ na nchekwa;Arụ ọrụ ịsacha PW buffer ezighi ezi, butere mfu RNase, na-emetụta ọrụ nnwale RNA nke ala, dị ka ntụgharị in vitro.
Nkwanye: Foregene usoro nucleic acid kewapụ ngwa nwere ike wepu RNA na-enweghị mgbakwunye RNase ọzọ, yabụ buccal Swab/FTA Kaadị DNA Isolation Kit ekwesịghị itinye RNase;jide n'aka na ị ga-agbaso ntuziaka maka kọlụm ịsacha Buffer PW, na enweghị ike ịhapụ usoro a.
4. Ethanol fọdụrụnụ.
Nyocha: Buffer WB emeghị centrifugation tube efu ka ọ sachara kọlụm dị ọcha.
Nkwanye: Mee ezi ọrụ centrifugation tube efu dị ka ntuziaka si dị.
5. Mmetọ adịghị ọcha ndị ọzọ.
Analysis: Echekwara samples ma ọ bụ pụrụ iche sample na-adịghị pretreated.
Nkwanye: Hazie ihe nlele ahụ nke ọma dị ka enyere ya ntụziaka.
