Mouse Genotyping Kit

Katsi Nhamba: HCR2021A

Ichi chigadzirwa kititi chakagadzirirwa kukurumidza kuzivikanwa mbeva genotypes, kusanganisira DNA crude extraction uye PCR amplification system.Chigadzirwa chacho chinogona kushandiswa kukwidziridzwa kwePCR zvakananga kubva pamuswe wembeva, nzeve, chigunwe uye mamwe matishu mushure mekutsemuka kwakapusa naLysis Buffer uye Proteinase k.Hapana kugaya kwehusiku, phenol-chloroform kudhirowa kana kucheneswa kwekoramu, iri nyore uye inopfupisa nguva yekuedza.Chigadzirwa chacho chakakodzera kukwidziridzwa kwezvimedu zvinonangwa kusvika ku2kb uye multiplex PCR maitiro ane anosvika matatu mapeya ekutanga.Iyo 2 × Mouse Tissue Direct PCR Mix ine genetically engineered DNA polymerase, Mg.²⁺, dNTPs uye yakagadziridzwa buffer sisitimu yekupa yakakwira amplification kunyatsoshanda uye inhibitor kushivirira, kuitira kuti PCR maitiro anogona kuitwa nekuwedzera template uye primers uye rehydrating chigadzirwa ku1 ×.Iyo PCR chigadzirwa chakakwidziridzwa nechigadzirwa ichi chine yakakurumbira "A" base kumagumo e3 uye inogona kushandiswa yakananga TA cloning mushure mekucheneswa.

Zvikamu

Storage Conditions

Zvigadzirwa zvinofanirwa kuchengetwa pa -25 ~ -15 ℃ kwemakore maviri.Mushure mekunyunguduka, Lysis Buffer inogona kuchengetwa pa2 ~ 8 ℃ yenguva pfupi-pfupi kushandiswa kwakawanda, uye sanganisa zvakanaka kana uchishandisa.

Application

Ichi chigadzirwa chakakodzera mbeva kugogodza kuongororwa, transgenic kutariswa, genotyping uye zvichingodaro.

Features

1.Kushanda kuri nyore: hapana chikonzero chekubvisa genomic DNA;

2.Wide application: yakakodzera kukwidziridzwa kwakananga kweakasiyana mbeva tishu.

Mirayiridzo

1.Kuburitswa kwegenomic DNA

1) Kugadzirira kwe lysate

Tissue lysate inogadzirirwa zvichienderana nehuwandu hwemasampunzi egonzo kuti aise lysed (tishu lysate inofanirwa kugadzirirwa pa-saiti zvinoenderana nedosi uye yakasanganiswa zvakakwana kuti ishandiswe), uye chikamu chemareagents chinodiwa kune imwechete sampu ndeichi:

2) Muenzaniso Kugadzirira uye Lysis

Inokurudzirwa Kushandisa Tissue

Tora huwandu hwakakodzera hwemasikirwo emakonzo mumachubhu ecentrifuge akachena, wedzera 200μL yetishu itsva lysate kune yega yega centrifuge chubhu, vortex uye zunza, wozoisa pa55 ℃ kwe30mins, wozopisa pa98℃ kwe3mins.

3) Centrifugation

Zunza iyo lysate zvakanaka uye centrifuge pa12,000 rpm kwe5mins.Iyo supernatant inogona kushandiswa setemplate yePCR amplification.Kana template ichidikanwa kuchengetedza, endesa iyo supernatant kune imwe isina centrifuge chubhu uye chengeta pa -20 ℃ kwemavhiki maviri.

2.PCR Amplification

Bvisa iyo 2 × Mouse Tissue Direct PCR Mix kubva -20 ℃ uye nyunguduka pachando, sanganisa kumusoro uye gadzirira PCR reaction system zvinoenderana netafura inotevera (shanda pachando):

Cherechedza:

a) Mari yakawedzerwa haifanirwe kudarika 1/10 yehurongwa, uye kana yakawandisa yawedzerwa, PCR amplification inogona kudziviswa.

Yakakurudzirwa PCR Conditions

Cherechedza:

a) Anealing tembiricha: Nekureva kukosha kweTm yeprimer, zvinokurudzirwa kuseta tembiricha yekumisa kune diki Tm kukosha kweprimer +3 ~ 5 ℃.

Matambudziko Anowanikwa uye Mhinduro

1.Hapana mitsetse yakanangwa

1) Yakawandisa lysis chigadzirwa.Sarudza huwandu hwakakodzera hwetemplate, kazhinji isingapfuuri 1/10 yehurongwa;

2) Saizi yakakura kwazvo.Dilute iyo lysate kagumi uye wobva wawedzera, kana kuderedza saizi yemuenzaniso uye re-lysis;

3) Tissue samples haisi nyowani.Inokurudzirwa kushandisa sampuli itsva yetishu;

4) Yakashata primer quality.Shandisa genomic DNA yekukwidziridza kuratidza iyo primer mhando uye optimize dhizaini yekutanga.

2.Non-specific amplification

1) Iyo annealing tembiricha yakadzikira uye nhamba yekutenderera yakakwira zvakanyanya.Wedzera tembiricha yekumisa uye kuderedza huwandu hwema cycle;

2) Template concentration yakanyanyisa.Deredza huwandu hwetemplate kana kuderedza template 10 nguva mushure mekusimudzira;

3) Yakashata primer chaiyo.Gadzirisa iyo yekutanga dhizaini.

Notes

1.Kuti udzivise kusvibiswa kwepakati pakati pemasampuli, maturusi akawanda ekuenzanisa anofanirwa kugadzirirwa, uye pamusoro pezvishandiso zvinogona kucheneswa ne2% sodium hypochlorite solution kana nucleic acid cleaner mushure meimwe sampling kana kushandiswa kudzokororwa kuchidiwa.

2.Zvinokurudzirwa kushandisa nyowani mbeva matishu, uye sampling vhoriyamu haifanirwe kunge yakakura zvakanyanya kudzivirira kukanganisa mhedzisiro yeamplification.

3.Lysis Buffer inofanirwa kudzivirira kazhinji kutonhora-kunyungudika, uye inogona kuchengetwa pa2 ~ 8 ℃ kwekushandisa kwenguva pfupi yakawanda.Kana yakachengetwa patembiricha yakaderera, kunaya kwemvura kunogona kuitika, uye inofanira kunyungudika zvizere isati yashandiswa.

4.PCR Musanganiswa inofanirwa kudzivirira kazhinji kutonhora-inonyungudika, uye inogona kuchengetwa pa4 ℃ kuitira kushandiswa kwenguva pfupi inodzokororwa.

5.Ichi chigadzirwa ndechekutsvagisa kwesainzi uye hachifanirwe kushandiswa mukuongororwa kwekiriniki kana kurapwa.