基因工程原理实验指导 3.The B-actin is used as the internal standard for each RT-PCR,DNA contamination in the RNA sample is tested by replacing the reverse transcriptase with water in the RT-PCR. Reagents 5x first strand buffer(GIBCO Part No.Y00146) 100 mM DTT (GIBCO Part No.Y11147) 200 U/uI M-MLV Reverse transcriptase(GIBCO Part No.28025-021) 500 ug/ml oligo(dT)15 primer(Promega Cat.No.C110A9362612) 10 mM dNTP mix RNase-free DNase I(TaKaRa Code No.2215 CA) 5 U/uI Ex Taq polymerase and 10 x buffer (TaKaRa Code No.RR001D CA) 10 HIM gene specific Primer F and R Sterile ddH2O Mineral oil 附录一细菌培养基 LB培养基 NaCl 10g Yeast extract 5g Peptone 10g Add dho to 1000ml Aliquot to 500ml flasks(250 ml per flask).Seal with sealfilm and autoclave them.Store at room temperature. LA固体培养基 NaCl 10g Yeast extract 5g Peptone 10g Agar powder 13~15g Add dH2O to 1000ml Aliquot to 500 ml flasks(250 ml per flask).Seal with sealfilm and autoclave them.Store at room temperature. X-gal (20mg/ml) 20mgX-gal溶于1ml二甲基甲酰胺中，-20℃避光保存： IPTG(200mg/ml): 1gPTG溶于4ml去离子双蒸水中，定容至5ml,过滤灭菌后20℃保存 3基因工程原理实验指导 21 3. The β -actin is used as the internal standard for each RT-PCR, DNA contamination in the RNA sample is tested by replacing the reverse transcriptase with water in the RT-PCR. Reagents 5 × first strand buffer (GIBCO Part No.Y00146) 100 mM DTT (GIBCO Part No.Y11147) 200 U / µ l M-MLV Reverse transcriptase (GIBCO Part No.28025-021) 500 µ g / ml oligo (dT) 15 primer (Promega Cat. No. C110A 9362612) 10 mM dNTP mix RNase-free DNase I (TaKaRa Code No. 2215 CA) 5 U / µ l Ex Taq polymerase and 10 × buffer (TaKaRa Code No. RR001D CA) 10 µ l M gene specific Primer F and R Sterile ddH 2 O Mineral oil 附录一 细菌培养基 LB 培养基 NaCl 10 g Yeast extract 5 g Peptone 10 g Add dH2O to 1000 ml Aliquot to 500ml flasks (250 ml per flask). Seal with sealfilm and autoclave them. Store at room temperature. LA 固体培养基 NaCl 10 g Yeast extract 5 g Peptone 10 g Agar powder 13 ～ 15 g Add dH2O to 1000 ml Aliquot to 500 ml flasks (250 ml per flask). Seal with sealfilm and autoclave them. Store at room temperature. X-gal (20mg/ml) ： 20mg X-gal 溶于 1ml 二甲基甲酰胺中， -20 ℃避光保存； IPTG (200mg/ml) ： 1g IPTG 溶于 4ml 去离子双蒸水中，定容至 5ml ，过滤灭菌后 -20 ℃保存；