2.2 mL, Open-Top Thinwall Ultr

Single tube confirmation PCR protocol

Polymerase. - Add 35 µl of the PCR mix to each of the 20 PCR tubes that already contain the 15 µl of primers and template. 110 µl 5 µl 10 x Taq buffer(see below) 11 µl 0.5 µl 20 mM dNTP's (0.2 mM) 11 µl 0.5 µ

Purification of Plasmid from 50 ml-culture

4. Resuspend the pellet in 5 ml of Solution I. 5. Add 10 ml of Solution II. Mix well by inverting the bottle more than 10 times. The solution should be clear after mixing Solution II. 6. Add 7.5 ml of Solution III. Mix well as above. 7. Spin

Transformation Ultra-Competent E. coli

4. Resuspend cells gently in 80 ml of ice cold " TB ". 5. On ice for 10 minutes. 6. Spin at 2500 x g (5000 rpm in a Sorvall GSA, 5500 rpm in a Sorvall SS-34, or 3000 rpm in a Beckman J-6B centrifuge) for 10 min. at 4C. 7. Resuspend cells gently in 20