Agrobacterium plasmid quick-screening

>>Agrobacterium cells grown from a colony in 5 ml of fresh LB medium (plus appropriate antibiotics) with vigorous stirring overnight at 280C in the dark).
>>The culture of transfer in the PE pipe and harvesting (centrifugation at 7000-8000 rpm. / Min after 10-15 seconds)
>>Resuspent cell pellets with 100 UL cold Sol I + 4 mg / ml lysozyme and incubate 30-40 minutes at room temperature.
>>Add 200 UL fresh Sol II to mix and shake vigorously (do not vortex) and incubated for 15 minutes at room temperature.
>>Add 30 ul phenol (balanced with 2 volumes of Sol II) and stir gently for several Seconde.
>>Add 150 UL 3M NaOAc (pH 4.8) and shake the tube briefly.
>>Keep the tube-200C for 20 minutes.
>>Centrifuge at 10000 rpm for 10 minutes and transfer supernatant to a new tube Parliament.
>>Add 1 volume of phenol (balanced) and mix and centrifuge at 14,000 rpm for 15 minutes.Download supenatant new EP.
>>Add 1 volume of chloroform and mix well and centrifuge at 14,000 ROM for 15 minutes at 40C.Download supenatant new EP.
>>Fill the tube with ethanol 100% vol 2 cold stored at-200C for 30 minutes.
>>Centrifuge for 15 minutes at 15,000 rpm. / Min 40 ° C and then discard the supernatant.
>>Wash with 70% ethanol and drying of the RT (dry or speed-vac).
>>Dissolve the DNA pellet with 20 ul DW.
Recipe:
Solution I
50 mM glucose
25 mm Tris-HCl (pH 8.0)
10 mm EDTA (pH 8.0)
DW 200 ml
Autoclave, store at 40 ° C

Decision II
0.2 N NaOH
1% (W / V) SDS
DW 200 ml
Store at RT

Sodium acetate 3M, pH 4.8
Phenol (balanced with 2 vol Sol II) (bacth ice)
Chloroform (bacth ice)
Ethanol 100% and 70% (cold)