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文献和实验Preparation of Microinjection Buffer
EDTA, pH 8.0 (autoclaved) 100 mM NaCl 1 ml of 5 M NaCl (autoclaved) 1x Polyamines 50 microliters 1000x Polyamines mix Autoclaved H2O up to 50 ml NOTE: Prepare fresh and discard unused microinjection buffer.
Phosphoamino acid analysis:Mark Kamps's method
of each buffer. pH 1.9 buffer. 88% Formic acid 50 ml Acetic acid 156 ml H2O 1794 ml Don't use the 98% formic acid and don't adjust the pH. pH 3.5 buffer Pyridine 10 ml Acetic acid 100 ml 100 mmolar EDTA 10 ml H2O 1880 ml Don't bother to adjust
CCMB buffer to make chemical competent cell
does not appear to be necessary Test competence (see below) Good cells should yield around 100 - 400 colonies 4. CCMB80 buffer 10 mM KOAc pH 7.0 (10 ml of a 1M stock/l) 80 mM CaCl2.2H2O (11.8 g/l) 20 mM MnCl2.4H2O (4.0 g/l) 10 mM MgCl2.6H2O (2.0 g/l) 10
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