pBR322 with rop: 10 - 25 copies/cell.
pBR322 without rop: 45 - 60 copies/cell.
pUC with rop: 10 - 25 copies/cell.
pUC without rop: 500 -700 copies/cell.

The high copy number of pUC plasmids is a result of the lack of the rop gene and a single point mutation in pMB1

Buffer P1 (resuspension buffer)
50 mM Tris·Cl, pH 8.0; 2–8°C, after
 10 mM EDTA; addition of
 100 μg/ml RNase A RNase A

Buffer P2 (lysis buffer)
200 mM NaOH, 1% SDS (w/v) 15–25°C

Buffer P3 (neutralization buffer)
 3.0 M potassium acetate,  pH 5.5

Buffer FWB2 (QIAfilter wash buffer)
1 M potassium acetate 15–25°C, pH 5.0

Buffer QBT (equilibration buffer)
 750 mM NaCl;
 50 mM MOPS, pH 7.0;
 15% isopropanol (v/v);
 0.15% Triton® X-100 (v/v)

Buffer QC (wash buffer)
 1.0 M NaCl
 50 mM MOPS, pH 7.0;
 15% isopropanol (v/v)

Buffer QF (elution buffer)
 1.25 M NaCl; 15–25°C
 50 mM Tris·Cl, pH 8.5;
 15% isopropanol (v/v)

Buffer QN (elution buffer)
  1.6 M NaCl; 
 50 mM MOPS, pH 7.0;
 15% isopropanol (v/v)

TE
 10 mM Tris·Cl, pH 8.0; 15–25°C
 1 mM EDTA

STE
 100 mM NaCl;
 10 mM Tris·Cl, pH 8.0;
 1 mM EDTA

P1:
Dissolve 6.06 g Tris base, 3.72 g Na2EDTA·2H2O in 800 ml distilled water.  Adjust the pH to 8.0 with HCl. Adjust the volume to 1 liter with distilled water.  Add 100 mg RNase A per liter of P1.

P2:
Dissolve 8.0 g NaOH pellets in 950 ml distilled water, 50 ml 20% SDS (w/v)    solution. The final volume should be 1 liter.

P3:
Dissolve 294.5 g potassium acetate in 500 ml distilled water. Adjust the pH to    5.5 with glacial acetic acid (~110 ml). Adjust the volume to 1 liter with distilled   water.

FWB2:
Dissolve 98.2 g potassium acetate in 500 ml distilled water. Adjust the pH to 5.0 with glacial acetic acid (~36 ml). Adjust the volume to 1 liter with distilled water.

QBT:
Dissolve 43.83 g NaCl, 10.46 g MOPS (free acid) in 800 ml distilled water. Adjust the pH to 7.0 with NaOH. Add 150 ml pure isopropanol and 15 ml 10% Triton X-100 solution (v/v). Adjust the volume to 1 liter with distilled water.

QC:
Dissolve 58.44 g NaCl and 10.46 g MOPS (free acid) in 800 ml distilled water.    Adjust the pH to 7.0 with NaOH. Add 150 ml pure isopropanol. Adjust the volume   to 1 liter with distilled water.

QF:

Dissolve 73.05 g NaCl and 6.06 g Tris base in 800 ml distilled water and adjust    the pH to 8.5 with HCl. Add 150 ml pure isopropanol. Adjust the volume to   1 liter with distilled water.

QN:
 Dissolve 93.50 g NaCl and 10.46 g MOPS (free acid) in 800 ml distilled water    and adjust the pH to 7.0 with NaOH. Add 150 ml pure isopropanol. Adjust the   volume to 1 liter with distilled water.

STE:
Dissolve 5.84 g NaCl, 1.21 g Tris base, and 0.37 g Na2EDTA·2H2O in 800 ml      distilled water. Adjust the pH to 8.0 with HCl. Adjust the volume to 1 liter with     distilled water

AcGFP: U.S. 7,432,053
DsRed-Monomer and Fruit Fluorescent Proteins: U.S. Patents: 7,157,566; 7,393,923; 7,005,511 and 7,250,298.
E2 Crimson: pending
GFP and GFPuv: U.S. Patents: 5,491,084 and/or 6,146,826,  5,605,793 and 5,811,238.
TagGFP, TagBFP, TagCFP, TagYFP, TagRFP, mKate etc: U.S. Patents: 7,417,131; 7,678,893; 7,605,230; 7,638,615, Evrogen
Monster Green Fluorescent Protein: U.S. Pat. Nos. 7,291,711 and 7,413,874
SuperFolder GFP

1, Culture cells using LB or 2xYT.
2, Spin down cells
3, Suspend cells in 10 ml of ice-cold P1 (50 mM Tris.HCl pH 8.0, 10 mM EDTA pH 8.0, 100 ug/ml RNase A).
4, Lyse cells with with 10 ml P2 (0.2 M NaOH, 1% SDS)
5, Add 10 ml P3 (3 M potassium acetate, pH 5.3 with acetic acid).
6, Spin at 8,000 g for 5 minutes at 4 oC .
7, Add 0.5 vol of  ice-cold 6 M guanidine hydrochloride in 10% acetic acid to cleared supernatant (final concentration of guanidine hydrochloride is 2 M).
8, Add mixture to syringe attached to 4 glass syringe filters in tandem.
9, Push plunger to make flow rate ~15 ml/min (total time is 2 min).
10, Wash filters with 20 ml PE (2 mM Tris.HCl pH 7.5, 20 mM NaCl in 80% EtOH).
11, Elute DNA with 10 ml of TE (pH 8.0).
12, Add 2 ml of 3 M sodium acetate (pH 5.1) and 15 ml of ice-cold isopropyl alcohol.
13, Put on ice for 5 minutes.
14, Load solution to syringe attached to a new set of 4 glass syringe filters in tandem.
15, Wash filters with 10 ml of 70% ethanol.
16, Elute DNA with 1 ml TE (pH 8.0) (incubation time with TE is ~3 min).

Materials:

1, 30 mm glass syringe filters with a 0.7 um pore-size: National Scientific Co. (cat #: F2500-18)
2, 25 mm glass syringe filters with a 1 um pore-size: Pall Life Science (cat #: 4523T).
3, 25 mm glass filter discs with a 0.7 um pore-size: Millipore (cat #: APFF02500);   filter holder: Sterlitech (cat #: 540100).

Capacity: 100 ug DNA per filter.

1, Add Triton X-114 to DNA, on ice, to a final concentration of 1% v/v.
2, Vortex at high speed 10 seconds.
3, Heat to 45°C for 5 minutes.
4, Centrifuge 5 minutes at room temp
5, Take supernatant (top layer) to new tube, chill on ice