Difference between revisions of "PhiX:Recipes"
From CCGB
| Line 21: | Line 21: | ||
dissolve in nuclease-free water and store at -20<sup>o</sup>C | dissolve in nuclease-free water and store at -20<sup>o</sup>C | ||
| − | ''' | + | '''10 X PE1'''<br> |
200mM Tris pH 7.5<br> | 200mM Tris pH 7.5<br> | ||
100mM MgCl<sub>2</sub><br> | 100mM MgCl<sub>2</sub><br> | ||
| Line 27: | Line 27: | ||
10 mM DTT | 10 mM DTT | ||
| − | ''' | + | '''10 X PE2'''<br> |
200 mM Tris pH 7.5<br> | 200 mM Tris pH 7.5<br> | ||
100 mM MgCl<sub>2</sub><br> | 100 mM MgCl<sub>2</sub><br> | ||
100 mM DTT | 100 mM DTT | ||
Revision as of 17:12, 18 August 2009
BE Buffer (one litre; 50mM Na2B4O7; 5.0mM EDTA)
19.1g Na2B4O7.10H2O
1.86g EDTA (disodium salt)
water to 1 litre
Tris-saturated Phenol with antioxidant (with 100ml phenol)
add 100mg 8-hydroxyquinoline (i.e., 0.1% w/v)
add equilibration buffer (-> pH 7.9) - 6.5ml of this in Sigma kit
add 130ul 2-mercaptoethanol (0.2% of buffer phase)
EDTA stock (0.5M)
use disodium salt and pH to 8 with NaOH
SDS 20%
pre-bought - avoids fine powder
Tris-HCl stock (1M)
pH to 8 with HCl
Pronase to 20mg/ml
dissolve in nuclease-free water and store at -20oC
10 X PE1
200mM Tris pH 7.5
100mM MgCl2
200 mM NaCl
10 mM DTT
10 X PE2
200 mM Tris pH 7.5
100 mM MgCl2
100 mM DTT
