KCTC Media No. 1509 : DCB-1 medium

조성	용량	단위
NaCl	1.0	g/L
MgCl2·6H2O	0.5	g/L
KH2PO4	0.2	g/L
NH4Cl	0.3	g/L
KCl	0.3	g/L
CaCl2·2H2O	0.015	g/L
100x salts	10.0	ml
trace element solution	1.0	ml
조성 용량 단위 HCl(25% solution, w/w) 10.0 ml FeCl2·4H2O 1.5 g CoCl2·6H2O 0.19 g MnCl2·4H2O 0.1 g ZnCl2 70 mg H3BO3 6 mg Na2MoO4·2H2O 36 mg NiCl2·6H2O 24 mg CuCl2·2H2O 2.0 mg First dissolve FeCl2·4H2O, 1.5g in HCl then dilute to water!!!)
Se/Wo solution	1.0	ml
조성 용량 단위 Na2SeO3·5H2O 6 mg/L NaOH 0.5 g/L Na2WO4·2H2O 8 mg/L
Resazurin(0.1% solution)	0.25	ml
5mM formate (electron donor)	0.34005	g/L
5mM acetate (carbon source)	0.41015	g/L
H2O bidest	add 1000	ml

** ph : 7.0-7.2

** memo : - some chemicals(e.g. fumarate, pyruvate) should not be autoclaved.
- boil, cool down to room temperature under fluching with N_2_/CO_2_(80/20)
- add 30mM NaHCO_3_
- adjust pH 7.0-7.2 with CO_2_(to lower the pH with CO_2_ is less time consuming)
- vary N_2_ and CO_2_ flow rates until stable desired pH is seached(discontinue stirring to decrease gas exchange)
- add reductant after stable pH is obtained CO_S_(3options):
1) add 0.2mM L-cysteine(MW 157.6) 0.031g/L
AND add 0.2mM Na_2_S*9H_2_O 0.048g/L
2) add 1mM DL-dithiothreitol 0.154g/L
- flush serum bottles with N_2_/CO_2_
- dispense medium(ca. 98ml per bottle), close the bottles with black rubber stoppers(or Teflon-lined stoppers)
- autoclave after medium turned clear; autoclave bottles in a closed basket; do NOT remove bottles from autoclave before temperature of the medium has reached<70℃

* Elctron donor(formate:5mM), acceptor(tetrachloroethene: 0.6mM), and carbon source(acetate: 5mM) can be added prior to heat sterilization.

**Attention:
The dissolution of tetrachloroethene requires ultrasound.

** 추가조치 사항 : -