2×PCR Super Mix (me ka Pelu)

2× PCR Master Mix loaʻa iā Taq DNA Polymerase, dNTPs, a me nā mea e pono ai ka PCR.Paʻa ka Master Mix no 3 mau mahina ma 4 ℃ me kā mākou mau stabilizer maʻamau.Hoʻopili ʻia ka hopena pre-mix no ka PCR maʻamau a mākaukau e hoʻohana me ka hoʻohui ʻana i ka DNA template a me nā primers.Hiki ke hoʻouka pololei ʻia nā huahana PCR no ka electrophoresis me ka wai kala uliuli bromophenol.Loaʻa i nā huahana i hoʻonui ʻia he 3 '-dA protrusion a hiki ke hoʻopili maʻalahi ʻia i T vector.Hoʻomaʻamaʻa ka 2 × PCR Master Mix i ke kaʻina hana PCR a hōʻemi i ka ʻino.

Nā Kūlana Waihona

Pono e mālama ʻia nā huahana ma -25 ℃ ~ -15 ℃ no 2 mau makahiki.

Nā kikoʻī

Nā kuhikuhi

1.Pūnaehana Manaʻo

2.Hoʻolaha Hoʻolaha

Nānā:

1) Ka hoʻohana ʻana i ka template: 50-200ng genomic DNA;0.1-10ng plasmid DNA.

2) Mg²⁺ka manaʻo: Loaʻa i kēia huahana he 3mM o MgCl2, kūpono no ka hapa nui o nā hopena PCR.

3) Ke ana wela: E ʻoluʻolu e nānā i ka waiwai Tm kumu o nā kumu mua.Hiki ke hoʻonohonoho ʻia ka mahana annealing i 2-5 ℃ haʻahaʻa ma mua o ka waiwai kumu o ka primer.

4) Ka manawa hoʻonui: No ka ʻike molekala, ʻōlelo ʻia ʻo 30 sec/kb.No ka hoʻopili ʻana i nā gene, ʻōlelo ʻia ʻo 60 sec/kb.

Nā memo

1.ʻAʻole kūpono nā huahana PCR me 2 × PCR Master Mix no ka polyacrylamide gel electrophoresis.

2.No kou palekana a me kou olakino, e ʻoluʻolu e ʻaʻahu i nā ʻaʻahu lab a me nā mīkina lima hoʻopau no ka hana.

3.Hoʻohana ʻia no ka noiʻi WALE!