Category: Protocols

Receipe for 10*TAE Buffer : 1Litre

                                                  10x           50x

1) Tris base                         48.4g          242g

2) Glacial acetic acid      11.42 ml    57.1ml

3) 0.5 EDTA                        20ml     100ml (PH 8.0)

    Total Volume                  1Liter       1L

Some of the simple techniques useful for students like me.

1. Overnight 5ml subculture at 37 ℃ for 18hr.
2. Inoculate 100µl subculture into 100 ml LB broth.
3. Culture at 37 ℃, 2½ hr.
4. Centrifuge at 4000 rpm, 10mins at 4°C (50ml +50ml).
5. Re-suspend cell pellet in 5ml 0.1 M cacl2 for each tube.
6. Keep on ice for 5mins.
7. Centrifuge.
8. Finally re-suspend cell pellet in 3 ml 0.1 m Cacl½ (1.5ml for each )
9. Preserve at -70 ℃ by adding 600µl of 50% glycerol ( final 10% gly) and aliquot into 1.5 ml tubes containing 200µl each.