PCR targeting system in Streptomyces coelicolor ... - Streptomyces UK

1 Make up a 1M solution of Tris and adjust to pH8.0 by adding maleic acid. For use,
add 3 parts by volume of the above solution to 1 part by weight of PEG 1000,
previously sterilised by autoclaving.
L (lysis) buffer (Thompson et al., 1982)
Mix the following sterile solutions:
Sucrose (10.3%) 100 ml
TES buffer (5.73%, adjusted to pH7.2) 10 ml
K2SO4 (2.5%) 1 ml
1
Trace element solution 0.2 ml
KH2PO4 (0.5%) 1 ml
MgCl2·6H2O (2.5M) 0.1 ml
CaCl2 (0.25M) 1 ml
1 The same as in R2 and R2YE.
This stock solution keeps indefinitely. Just before use dissolve lysozyme in a sample
of the solution at a concentration of 1 mg/ml and sterilise by filtration.
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