Transformation of competent E.coli cells via the heat-shock method.
Reagents:
LB Broth (1L):
Tryptone - 10g
Yeast Extract - 5g
Sodium Chloride - 10g
Mix, add DI H2O, autoclave, cool and use or keep in sealed autoclaved containers until needed.
LB Agar (1L):
Peptone -10g
Yeast Extract - 5g
Sodium Chloride - 10g
Agar - 12g
Dissolve in DI H2O, autoclave, wait until cool to the touch, add proper concentration of antibiotic if needed, pour 10-20ml per plate, allow agar to cool and harden. Use, or seal and keep at +4 until needed
Protocol:
1. Thaw cells and DNA on ice (15-20min).
2. Mix DNA with cells, incubate on ice 15 min.
3. Heat shock at 42° C for 2 min.
4. Cool on ice for 3 min.
5. add 100ul LB, shake for 1 hour at 37° C.
6. Spread on plate
7. Place in incubator overnight at 37° C
Any questions do not hesitate to ask. It is better to ask than waste reagents doing blind experiments
