Erratum: Rapid Analysis of Chromosome Aberrations in Mouse B Lymphocytes by PNA-FISH

Riepilogo

A correction was made to: Rapid Analysis of Chromosome Aberrations in Mouse B Lymphocytes by PNA-FISH.

Abstract

A correction was made to: Rapid Analysis of Chromosome Aberrations in Mouse B Lymphocytes by PNA-FISH. Table 1 (the list of solutions to prepare) was omitted from the Protocol section. The contents of Table 1 are as follows:

Solution	Reagents	Comments	Storage
B Cell Wash Buffer	500 ml 1x Hanks Balanced Salt Solution	Filter through 0.22 μm stericup filtration unit	4 °C
	5 ml 50 °C heat inactivated FCS
	5 ml 100x Pen/Strep
B Cell Medium	418.5 ml RPMI-1640	Filter through 0.22 μm stericup filtration unit	4 °C
	50 ml 50 °C heat inactivated FCS
	5 ml 100x Pen/Strep
	5 ml glutamine
	5 ml nonessential amino acids
	5 ml sodium pyruvate
	1.8 ml 14.2 M 2-mercaptoethanol
	5 ml 1M HEPES
Fixative	3:1 v/v methanol/glacial acetic acid		Make fresh
0.075 M KCl	1.395 g KCl		Room Temperature
	250 ml distilled H2O
FISH Master Mix	40 ml 50% Dextran sulfate	Vortex the solution and leave in a shaking platform overnight	Aliquot and store at -20 °C
	20 ml 20x SSC
	40 ml sterile distilled H2O
70% Formamide/2x SSC	10 ml 20x SSC	Adjust to pH 7 with 1M HCl	Aliquot and store at -20 °C
	20 ml distilled H2O
	70 ml deionized formamide
0.01 M HCl	1 ml 1M HCl	Adjust to pH 2.0	Room Temperature
	99 ml distilled H2O
1x PBS/MgCl2	50 ml 1M MgCl2		Room Temperature
	950 ml 1x PBS
50% Formamide/2x SSC	20 ml 20x SSC	Adjust to pH 7.25 Formamide used in this step does not have to be deionized.	Make fresh
	80 ml distilled H2O
	100 ml formamide
4x SSC/0.1% Tween20	50 ml 20x SSC		Room Temperature
	200 ml distilled H2O
	250 μl Tween20
DAPI staining solution (80 ng/ml)	40 μl 0.2mg/ml DAPI stock		4 °C in light protected staining jar
	100 ml 2x SSC
Mowiol Mounting Medium	6 g glycerol	Incubate at 50 °C with stirring for 2hrs	Aliquot and store at 4 °C
	2.4 g Mowiol 4-88
	6 ml distilled H2O
	12 ml 0.2 M TrisCl pH 8.5
	230 μl 1% Thimerosal (w/v in water)

Table 1. List of solutions. 

Protocollo

A correction was made to: Rapid Analysis of Chromosome Aberrations in Mouse B Lymphocytes by PNA-FISH. Table 1 (the list of solutions to prepare) was omitted from the Protocol section. The contents of Table 1 are as follows:

Solution	Reagents	Comments	Storage
B Cell Wash Buffer	500 ml 1x Hanks Balanced Salt Solution	Filter through 0.22 μm stericup filtration unit	4 °C
	5 ml 50 °C heat inactivated FCS
	5 ml 100x Pen/Strep
B Cell Medium	418.5 ml RPMI-1640	Filter through 0.22 μm stericup filtration unit	4 °C
	50 ml 50 °C heat inactivated FCS
	5 ml 100x Pen/Strep
	5 ml glutamine
	5 ml nonessential amino acids
	5 ml sodium pyruvate
	1.8 ml 14.2 M 2-mercaptoethanol
	5 ml 1M HEPES
Fixative	3:1 v/v methanol/glacial acetic acid		Make fresh
0.075 M KCl	1.395 g KCl		Room Temperature
	250 ml distilled H2O
FISH Master Mix	40 ml 50% Dextran sulfate	Vortex the solution and leave in a shaking platform overnight	Aliquot and store at -20 °C
	20 ml 20x SSC
	40 ml sterile distilled H2O
70% Formamide/2x SSC	10 ml 20x SSC	Adjust to pH 7 with 1M HCl	Aliquot and store at -20 °C
	20 ml distilled H2O
	70 ml deionized formamide
0.01 M HCl	1 ml 1M HCl	Adjust to pH 2.0	Room Temperature
	99 ml distilled H2O
1x PBS/MgCl2	50 ml 1M MgCl2		Room Temperature
	950 ml 1x PBS
50% Formamide/2x SSC	20 ml 20x SSC	Adjust to pH 7.25 Formamide used in this step does not have to be deionized.	Make fresh
	80 ml distilled H2O
	100 ml formamide
4x SSC/0.1% Tween20	50 ml 20x SSC		Room Temperature
	200 ml distilled H2O
	250 μl Tween20
DAPI staining solution (80 ng/ml)	40 μl 0.2mg/ml DAPI stock		4 °C in light protected staining jar
	100 ml 2x SSC
Mowiol Mounting Medium	6 g glycerol	Incubate at 50 °C with stirring for 2hrs	Aliquot and store at 4 °C
	2.4 g Mowiol 4-88
	6 ml distilled H2O
	12 ml 0.2 M TrisCl pH 8.5
	230 μl 1% Thimerosal (w/v in water)

Table 1. List of solutions.