Ropa Rakananga PCR Kit

Kukurumidza kukwidziridzwa kweiyo yakanangwa geni zvakananga kushandisa ropa setemplate isina kuburitswa.

Iyi kititi inotora genetically engineered anti-inhibitor DNA polymerase kuti ikwanise kuwedzera imwe-kopi majeni mune genome revanhu. Iyo yakanyatsogadziriswa buffer system mune ino kit inobatsira iyo polymerase yakasimba kudzivisa kuvharirwa kwePCR inhibitors, nokudaro inogona kunongedza zvakananga DNA ichishandisa ropa uye maseru akagadzirwa sematemplate. Ichi chigadzirwa chiri nyore kushanda uye hachidi matanho akaomarara senge DNA kuchenesa kana sampuro prereatment.
Iyi kit inopihwa se2 × MasterMix, uye maitiro acho anogona kuitwa nekungowedzera iyo template yeropa uye zvinoenderana nekuona kwekutanga. Inogona kuiswa pane akakwenenzverwa maseru emhuka dzakadai sevanhu, mbeva, nguruve, mombe uye mamwe marudzi, pamwe nerutsva kana 4 ℃ yakachengetedzwa yakazara ropa, anticoagulant (EDTA, citrate, heparin), yakanyorova ropa clots uye yakaoma nzvimbo dzeropa. zvakachengetwa pa Whatman 903 uye FTA Elute makadhi ekutengesa.

Cat. Aihwa	Kurongedza Saizi
4992529	20 ×l × 100 rxn
4992530	20 ×l × 500 rxn

Tumira neindaneti kwatiri

Chigadzirwa Chigadzirwa

Muenzaniso Wekuedza

MIBVUNZO

Chigadzirwa Mateki

Zvimiro

■ Zviri nyore uye zvinokurumidza: PCR kukwidziridzwa kunogona kuitwa zvakananga uchishandisa ropa setemplate, pasina kudikanwa kwematanho anonetesa ekuenzanisira gadziriro uye kubviswa kweDNA.
■ Kuchena kwepamusoro: Kusvetuka kwesampuro pre-kurapwa uye matanho ekubvisa eDNA anogona kubatsira kudzivirira kusvibiswa kwemasampuli.
■ Kubudirira kwepamusoro: Kuzivikanwa kwePCR kwemasampuru makuru kunogona kuitwa nekubatanidza kit pamwe nemaplate ePCR 96/384-tsime.
■ Simba rakasarudzika: Iyi kititi inogona kusimbisa zvakaringana zvidimbu zvepamusoro zveGC kana zvidimbu zvine chimiro chechipiri chakaomarara, uye kureba kwekukwiridzira kunogona kusvika pa5 kb.
Str Kusasimba kwekushushikana mupfungwa: Kitsi iyi inogona kuiswa kune akasiyana marudzi esampuli neropa zvakachengetedzwa nenzira dzakasiyana.

Zvishandiso

Iyo PCR zvigadzirwa zveichi kit zvine "A" pane 3'-kumagumo, iyo inogona kushandiswa zvakananga kune TA vector cloning. Ichi kit chinogona kushandiswa kukwidziridzwa kwe genomic DNA zvidimbu, yakakwira-kuburitsa mageneti ongororo uye genotyping (senge kuongorora kwegene) kuongorora.

Zvese zvigadzirwa zvinogona kuve zvakagadziriswa ODM / OEM. Kuti uwane mamwe mashoko,ndokumbira ubaye Customized Service (ODM / OEM)

Pashure: TIANcombi DNA Lyse & Det PCR Kit

Zvadaro: Mouse Tissue Yakananga PCR Kit

	Uchishandisa hwevanhu EDTA anticoagulation se template, magene mana ane akasiyana GC zvirimo akawedzeredzwa neRopa Direct PCR Kit. Iyo PCR reaction system yaive 20 μl, uye 1 μl ropa raishandiswa se template. M: TIANGEN Maki II; 1: Chidimbu saizi 1090 bp, GC zvemukati 68.1%; 2: Chidimbu saizi 1915 bp, GC zvemukati 70.4%; 3: Chidimbu saizi 448 bp, GC zvemukati 74.8%; 4: Chidimbu saizi 1527 bp, GC zvemukati 61.5%. Mhedzisiro yekuongorora: Ropa Rakanangana PCR Kit inogona kunyatsokwenenzvera zvidimbu zveDNA neGC zvemukati pane huwandu hwe6.5% -74.8%, zvichiratidza kuti inokwanisa kukwidziridza zvidimbu zvepamusoro-GC.
	Uchishandisa hwevanhu EDTA anticoagulation se template, mashanu majini ane urefu hwakasiyana (ActB, Prp, DN1.0, Hn2.0 uye Hn4.0) akawedzerwa neRopa Direct PCR Kit. Iyo PCR reaction system yaive 20 μl, uye 1 μl ropa raishandiswa se template. M: TIANGEN Maki II; 1-3: 3 dzakasiyana sampuli dzeropa; NTC: kutonga pasina primers. Mhedzisiro yekuongorora: Ropa Rakanangana PCR Kit inogona kukwidziridza zvidimbu nehurefu sekureba se4 kb, zvichiratidza kuti inokwanisa kukwidziridza zvidimbu zvakareba.
	Uchishandisa hwevanhu EDTA anticoagulation se template, Ropa Direct PCR Kit yakashandiswa pakuona kwePCR kwemasampula eropa akasiyana. Iyo PCR reaction system yaive 20 μl, uye 1 μl ropa raishandiswa se template. M: TIANGEN Maki II; 1-9: iyo yekurodha huwandu hweropa iri 0.1 μl, 0.2 μl, 0.3 μl, 0.4 μl, 1 μl, 2 μl, 3 μl, 4 μl uye 5 μl, zvichiteerana; NTC: kutonga pasina template Mhedzisiro yekuongorora: Ropa Rakanangana PCR Kit ine yakasimba kuramba muropa uye inogona kukwidziridza masampuli eropa pamwe neyekuregedza renji ye01-5 μl.
	Zviratidzo zveropa kubva kuvanhu, gonzo, huku uye dzimwe mhando dzine marapirwo akasiyana dzakashandiswa sematemplate. Ropa Direct PCR Kit yakashandiswa kukwidziridza PRNP (yevanhu, 750 bp), Actin (rat, 200 bp), uye β-Actin (huku, 1.0 kb). Iyo PCR reaction system yaive 20 μl, uye 1 μl ropa raishandiswa se template. M: TIANGEN Maki II. Mhedzisiro yekuongorora: Ropa Rakanangana PCR Kit inogona kuiswa pane akasiyana masampuli, uye iyo yakatarwa PCR kuona inogona kuitwa pamasampuli eropa kubva kune akasiyana marudzi ane akasiyana marapirwo.

Mubvunzo: Hapana mabhendi ekusimudzira

A-1 Chidimbu

■ Iyi template ine mapuroteni marara kana kuti Taq inhibitors, nezvimwe- ——Chenesa DNA template, bvisa tsvina yeprotein kana bvisa template yeDNA nemidziyo yekuchenesa.

■ Kuburitswa kwetemplate hakuna kukwana - - Zvakakodzera kuwedzera tembiricha yedenature uye kuwedzera nguva yekushandurwa.

■ Kushatiswa kwetemplate ——Gadzirisazve template.

A-2 Primer

■ Hunhu hwemhando dzekutanga - - Re-gadzira iyo yekutanga.

Prim Kushatiswa kwekutanga --—Rovera mabara epamusoro epamusoro kuva vhoriyamu diki yekuchengetedza. Dzivisa kuwanda kwechando uye kunyungudika kana kwenguva refu -4 ° C yakachengetedzwa.

■ Dhizaini isiriyo yezvigadzirwa (semuenzaniso, urefu hwepamberi haukwanire, dhayera yakaumbwa pakati pezvakagadzirwa, nezvimwewo) -Reresign primers (dzivisa kuumbwa kweprimer dimer uye yechipiri chimiro)

A-3 Mg²⁺kuisa pfungwa

■ Mg²⁺ kuiswa kwakanyanya kwazvo ——Kuwedzera zvakafanira Mg²⁺kusangana: Gonesa iyo Mg²⁺ kusunganidzwa neakateedzana ezvekuita kubva pa1 mM kusvika pa3 mM ine chinguva che0.5 mM kuona iyo yakanakisa Mg²⁺ kusungirirwa kwetemplate imwe neimwe uye yekutanga.

A-4 Kuwedzera kwekushisa

The Iyo tembiricha yekukwirisa inokanganisa kusunga kwekutanga uye template. - -Deredza iyo yekuwedzera tembiricha uye gadzirisa mamiriro acho ne gradient ye2 ° C.

A-5 Yekuwedzera nguva

■ Pfupi yekuwedzeredza nguva - - Wedzera nguva yekuwedzera.

Mubvunzo: Nhema dzakanaka

Phenomena: Negative sampuli zvakare inoratidza iwo akanangana akateedzana mabhendi.

A-1 Kusvibisa PCR

■ kusvibiswa kwemuchinjikwa kwekutevedzana kwekutarisa kana zvigadzirwa zvekusimudzira - - Nekuchenjera kusapomba sampuro ine nhevedzano yechinangwa mune yakashata sampuro kana kuvadururira kunze kwecentrifuge chubhu. Iwo ma reagents kana zvishandiso zvinofanirwa kuve zvakagadziriswa kuti zvibvise zviripo nucleic acids, uye kuvepo kwehutachiona kunofanirwa kutemerwa kuburikidza neasina kudzora kuyedza.

■ Kusvibisa reagent ——Root the reagents and store at low low.

A-2 Primer

■ Mg²⁺ kuiswa kwakanyanya kwazvo ——Kuwedzera zvakafanira Mg²⁺ kusangana: Gonesa iyo Mg²⁺ kusunganidzwa neakateedzana ezvekuita kubva pa1 mM kusvika pa3 mM ine chinguva che0.5 mM kuona iyo yakanakisa Mg²⁺ kusungirirwa kwetemplate imwe neimwe uye yekutanga.

■ Dhizaini isina kukodzera, uye maratidziro akanangwa ane humology pamwe neasingaitane nhevedzano. —-Re-dhizaini yekutanga.

Mubvunzo: Asina-chaiwo kududzira

Phenomena: Iwo maPC ekukudziridza mabhendi haapindirane nehukuru hunotarisirwa, ingave hombe kana diki, kana dzimwe nguva zvese zvakasarudzika mabhendi ekuwedzera uye asiri-akasarudzika mabhendi ekusimudzira anoitika.

A-1 Primer

■ Kusanyatsojeka kwekutanga

—-Re-dhizaini yekutanga.

■ Iko kusungirwa kwepamusorosoro kwakanyanyisa - - Zvakakodzera kuwedzera tembiricha yedenature uye kuwedzeredza nguva yekushanduka.

A-2 Mg²⁺ kuisa pfungwa

■ Iye Mg²⁺ kusungwa kwakanyanya ——Deredza zvakanaka iyo Mg2 + kukosheswa: Gonesa iyo Mg²⁺ kusunganidzwa neakateedzana ezvekuita kubva pa1 mM kusvika pa3 mM ine chinguva che0.5 mM kuona iyo yakanakisa Mg²⁺ kusungirirwa kwetemplate imwe neimwe uye yekutanga.

A-3 Thermostable polymerase

■ Huwandu hwe enzyme yakawandisa ——Deredza huwandu hwee enzyme nenzira kwayo panguva dze 0.5 U.

A-4 Kuwedzera kwekushisa

■ Tembiricha yekuvharisa yakanyanya kushomeka - - Zvakakodzera kuwedzera tembiricha yekuwedzera kana kutora nzira mbiri yekuvharisa

A-5 PCR kutenderera

■ Makwikwi mazhinji ePDR ——Deredza huwandu hwePCR macircule.

Mubvunzo: Patchy kana smear mabhendi

A-1 Primer-—Kusarongeka kwehurombo--- Gadzirisazve iyo yekutanga, chinja chinzvimbo uye kureba kweiyo yekutanga kuti iwedzere kujekesa kwayo; kana kuita PCR nested.

A-2 template DNA

——Template haina kuchena— - Chenesa template kana bvisa DNA nemidziyo yekuchenesa.

A-3 Mg²⁺ kuisa pfungwa

——Mg²⁺ kuisa pfungwa kwakanyanya ——Deredza zvakanaka Mg²⁺ kusangana: Gonesa iyo Mg²⁺ kusunganidzwa neakateedzana ezvekuita kubva pa1 mM kusvika pa3 mM ine chinguva che0.5 mM kuona iyo yakanakisa Mg²⁺ kusungirirwa kwetemplate imwe neimwe uye yekutanga.

A-4 dNTP

——Kusungwa kweDNTPs kwakanyanyisa ——Deredza kusangana kweDNTP nenzira kwayo

A-5 Kuwedzera tembiricha

——Tembiricha yakadzikira chose inovhara —— Zvishoma nemazvo wedzera tembiricha yekuwedzera

A-6 Matunhu

--—Kwakawanda kutenderera —— Wedzera huwandu hwenhamba

Mubvunzo: Mari yakawanda sei template inofanirwa kuwedzerwa mune 50 μl PCR reaction system?

Mubvunzo: Ungawedzera sei zvidimbu zvakareba?

Nhanho yekutanga ndeyekusarudza yakakodzera polymerase. Yakajairwa Taq polymerase haigone kuongorora nekuda kwekushayikwa kwe3'-5 'exonuclease chiitiko, uye kusaenderana kuchadzora zvakanyanya kuwedzera kwekuwedzera kwezvimedu. Naizvozvo, yenguva dzose Taq polymerase haigone kunyatsokwenenzvera zvidimbu zvakanangwa zvakakura kupfuura 5 kb. Taq polymerase ine yakakosha nadzurudzo kana imwe yakanyanya kutendeka polymerase inofanirwa kusarudzwa kuvandudza kuwedzera kwekuita uye kuzadzikisa zvido zvehurefu hwakareba kuwedzera. Uye zvakare, kukwidziridzwa kwezvimedu zvidimbu kunodawo kuenderana kunoenderana kwekutanga dhizaini, nguva yekuwedzeredza, nguva yekuwedzera, buffer pH, nezvimwe. Kazhinji, ma primers ane 18-24 bp anogona kutungamira mukukohwa kuri nani. Kuti tidzivirire kukuvara kwetemplate, iyo nguva yekuratidzira iri pa94 ° C inofanirwa kudzikiswa kusvika pamasekondi makumi matatu kana mashoma pasiringi, uye iyo nguva yekukwira tembiricha kusvika pa94 ° C pamberi pekukudziridza inofanira kunge iri pasi pe1 min. Zvakare, kumisikidza iyo yekuwedzera tembiricha ingangoita se68 ° C uye nekugadzira iyo yekuwedzera nguva zvinoenderana nehuremu hwe1 kb / min inogona kuve nechokwadi chinoshanda kukwidziridzwa kwezvimedu zvidimbu.

Mubvunzo: Ungavandudza sei kukwidziridzwa kuvimbika kwePCR?

Chiyero chekukanganisa kwePCR kukwidziridzwa kunogona kudzikiswa nekushandisa akasiyana maDNA ma polymerase nekuvimbika kwakanyanya. Pakati pese maTaq DNA ma polymerases akawanikwa kusvika parizvino, Pfu enzyme ine yakaderera kukanganisa chikanganiso uye yakatendeka kuvimbika (ona tafura yakasungirirwa). Pamusoro pekusarudzwa kwe enzyme, vatsvagiri vanogona kuwedzera kudzikisa huwandu hwePCR nekushandura mamiriro ekugadzirisa, kusanganisira kugadziridza bhaisikopo, kuwanda kweinopisa polymerase uye kugadzirisa huwandu hwePCR.

Nyora yako meseji pano uye uitumire kwatiri