Re: 哪位给贴个用PCR检查阳性克隆的protocol吧# Biology - 生物学
t*s
1 楼
1. Prepare PCR tubes with 5ul water in each;
2. prepare 15ml falcon tubes with 1ml of LB + antibiotics;
3. pick a colony using a sterile tip; stir it in a the water
-containing PCR tube for several times and then throw the
tip in the falcon tube to inoculate the LB medium.
4. use a primer on the vector and a primer on your insert
to prepare 15ul master mix for each 20ul reaction.
By using this primer set, you will be able to confirm
both availibility of insert and its orientation.
2. prepare 15ml falcon tubes with 1ml of LB + antibiotics;
3. pick a colony using a sterile tip; stir it in a the water
-containing PCR tube for several times and then throw the
tip in the falcon tube to inoculate the LB medium.
4. use a primer on the vector and a primer on your insert
to prepare 15ul master mix for each 20ul reaction.
By using this primer set, you will be able to confirm
both availibility of insert and its orientation.