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'''HIGH-SPEED GEL ELECTROPHORESIS_'''<br>
'''SB Buffer'''<br>
45g boric acid<br>
8g sodium hydroxide pellets<br>
water to 1 litre


'''PHAGE ELUTION_'''<br>
'''BE Buffer (50mM Na<sub>2</sub>B<sub>4</sub>O<sub>7</sub>; 5.0mM EDTA)'''<br>
19.1g Na<sub>2</sub>B<sub>4</sub>O<sub>7</sub>.10H<sub>2</sub>O<br>
1.86g EDTA (disodium salt)<br>
water to 1 litre


'''PHAGE GROWTH_'''<br>
'''LB Buffer'''<br>
10g tryptone<br>
5g yeast extract<br>
10g NaCl<br>
water to 1 litre, mix and autoclave<br>
after autoclaving add CaCl<sub>2</sub> to 2mM (2ml 1M per litre) and MgCl<sub>2</sub> to 10mM (10ml 1M per litre)

''For plate agar:''
add 15g Bacto brand agar per litre;
no need to add salts

''For top agar:''
add 7g Bacto brand agar per litre;
after autoclaving add CaCl<sub>2</sub> (to 2mM) and MgCl<2> (to 10mM)

''Note:''
Bentley Fane add CaCl<sub>2</sub> to 5mM


'''PHAGE STORAGE/DILUTION_'''<br>
'''HFB Buffer (60mM NH<sub>4</sub>Cl; 90mM NaCl; 0.1M KCl; 0.1M Tris-HCl (pH 7.4); 1.0mM MgSO<sub>4</sub>; 1.0mM CaCl<sub>2</sub>)'''<br>
''For 500ml 10× stock:''<br>
125ml of 2.0 M Tris-HCl (pH 7.3) -- surely 250ml?<br>
15g NH<sub>4</sub>Cl<br>
2.5g NaCl<br>
25g KCl<br>
5.0ml 1.0 M MgSO<sub>4</sub><br>
5.0ml 1.0M CaCl<sub>2</sub><br>
water up to 500ml<br>

''For working solution:''
dilute to 1× in water and autoclave


'''NUCLEIC ACID SEPARATION_'''<br>
'''Tris-saturated Phenol with antioxidant (with 100ml phenol)'''<br>
add 100mg 8-hydroxyquinoline (i.e., 0.1% w/v)<br>
add equilibration buffer (-> pH 7.9) - 6.5ml of this in Sigma kit<br>
add 130ul 2-mercaptoethanol (0.2% of buffer phase)


'''COMPONENT SOLUTIONS_'''<br>
'''EDTA stock (0.5M)'''<br>
use disodium salt and pH to 8 with NaOH

'''SDS 20%'''<br>
pre-bought - avoids fine powder

'''Tris-HCl stock (1M)'''<br>
pH to 8 with HCl

'''Pronase to 20mg/ml'''<br>
dissolve in nuclease-free water and store at -20<sup>o</sup>C

'''10 X PE1'''<br>
200mM Tris pH 7.5<br>
100mM MgCl<sub>2</sub><br>
200 mM NaCl<br>
10 mM DTT

'''10 X PE2'''<br>
200 mM Tris pH 7.5<br>
100 mM MgCl<sub>2</sub><br>
100 mM DTT