pHDEP 100/10/18*

(H2O)3.98

*

FON() (H+)FON (bonded pair : O-H)(lone pair:)) lone pair-lone pair> lone pair-bonded pair> bonded pair-bonded pair - *

*

*

1000/18 = 55.56 M)*

NaClKCl CH3COOHCH3COO-+H+*

1 NaCl Na+ + Cl -

2 ()*

(1) (2) (3)*

saline solution 0.670.700.850.9 6

()

12 ()()*2 H+ + 2 e- H2, H2O O2 + 2 H+ + 2e-

()*Semipermeable membraneSelectively permeable membranemoleculehttp://zh.wikipedia.org/wiki/%E5%8D%8A%E9%80%8F%E8%86%9C

(Vant Hoff) : =MRT (M=R= T=(K))M

Mhttp://zh.wikipedia.org*: i2

http://www.ccjhs.tpc.edu.tw*

(V)(I)V=I*R(R)

1 cmK1 cm-10.01 D KCl1408 S/cm 0.5 %, 25 ) =G*K=(1/R) *(L / A) http://www.ccjhs.tpc.edu.twK(S/cm)GL(cm)A(cm2)*

(HA)(A-)(BOH)(B)pH()()

HA+A-A-HABOH+B+BOH B+

CH3COOH + CH3COONa () NH4OH + NH4Cl () *

*pHpHpH

H2CO3HCO3- H+(aq) + HCO3-(aq) H2CO3 OH-(aq) + H2CO3(aq) H2O(l) + HCO3-(aq) pH7.2~7.4

*

(Phosphate)pH 6.5~7.5 (H3PO4)http://zhidao.baidu.com/question/54554006http://www.ihcworld.com/_protocols/washing_buffers/pb.htmPhosphate Buffer Saline (1 X PBS, pH 7.4) Na2HPO42H2O ----------------------- 1.4 g KH2PO4 ----------------------------------- 0.2 g NaCl ----------------------------------------- 8 g KCl ---------------------------------------- 0.2 g Distilled water ----------------------- 1000 ml Mix to dissolve and adjust pH to 7.4Phosphate Buffer (0.1 M PB, pH 7.4) Na2HPO4 (anhydrous) ----- 10.9 g NaH2PO4 (anhydrous) ----- - 3.2 g Distilled water --------------- 1000 ml Mix to dissolve and adjust pH to 7.4 Store this solution at room temperature*

Tris [Tris(hydroxymethyl)aminomethane]An abbreviation of the organic compoundTris is extensively used in biochemistry and molecular biology. In biochemistry, tris is widely used as a component of buffer solutions, such as in TAE and TBE buffer (involving nucleic acids, the most common being gel electrophoresis). Tris has a pKa of 8.06, pH range between 7.0 and 9.2 coincides with the typical physiological pH of most living organisms. This, and its low cost, make tris one of the most common buffers used in the biology/biochemistry lab.It is reported that Tris inhibits a number of enzymes. It should be used with care when studying proteins.C4H11NO3http://en.wikipedia.org/wiki/Tris*Amino group

Containing a mixture of Tris base, acetic acid and EDTA.It is made up of Tris-acetate buffer, usually at pH 8.0, and EDTA, which sequesters divalent cations. TAE has a lower buffer capacity than TBE and can easily become exhausted, but linear, double stranded DNA runs faster in TAE.TAE has been used to study the mobility of DNA in solution with and without sodium chloride (and many other salts). The salts will retards DNA mobility.TAE buffer offers advantages in subsequent enzymatic applications for the DNA sample. Borate in the TBE buffer is a strong inhibitor for many enzymes (DNA can better keep its integrity).*TAE buffer

TAE buffer& TBE bufferA mixture of Tris base, Acetic acid and EDTA(TAE) adjusted by NaOH, and the mixture of TBE is combined by Tris base, Boric acid and EDTA.

Tris-acid solutions are effective buffers for slightly basic conditions, which keep DNA deprotonated and soluble in water.

EDTA is a chelator of divalent cations, particularly of magnesium (Mg2+). As these ions are necessary co-factors for many enzymes, including contaminant nucleases, the role of the EDTA is to protect the nucleic acids against enzymatic degradation. But since Mg2+ is also a co-factor for many useful DNA-modifying enzymes such as restriction enzymes and DNA polymerases.*

pHMcIlvainepH *McIlvainepH2.6-12(Carmody) 1931-(Britton-Robinson)

(Carmody buffer)The Carmody buffer is a universal buffer composed of (x-y) ml of the basal buffer (containing 0.2 M boric acid and 0.05 M citric acid) and y ml of 0.1 M Na2PO412H2O and pure water as the pH adjuster, where x denotes the total volume of the buffer. It is very useful because it can be adjusted within a wide pH range (pH 2-12) by changing their ratios.*http://www.rsc.org/delivery/_ArticleLinking/DisplayArticleForFree.cfm?doi=b507540h&JournalCode=OBhttp://www.sciencedirect.com/science?_ob=MImg&_imagekey=B6T21-3XVPGV7-J-7&_cdi=4905&_user=9424682&_pii=S0167483899002162&_origin=search&_coverDate=11%2F16%2F1999&_sk=985649998&view=c&wchp=dGLbVlWzSkWb&md5=b045b3de417247f2134dad428b7f8a09&ie=/sdarticle.pdf

Goods Buffer - Composition and pH rangeGood's buffers are twelve buffering agents selected and described by Norman Good and colleagues in 1966. Good selected the buffers based on a number of criteria which make them candidates for use in biochemistry and biological research. Many remain staples in modern biology laboratories.http://www.tcieurope.eu/fr/product/bio-chem/B013.shtml

Buffer CapacitypH

CB and CA represent the concentrations of the added base "B" and acid "A".Buffer capacity is always positive. It is a non-lineal continuous function (the reciprocal of the titration curve slope). *http://www.aqa.org.ar/pdf9701/9701art16.pdf

pH pH = log10[H+] [H+]

pOHpHpOH = log10[OH]

25ion product of waterKw=[H+][OH-]=1.0110-14 .[H+]=[OH-]=1.00510-7 .pHwater=-log[H+]=-log1.00510-7=7 *

,, ,,, exBME( Basal Medium Eagle )Eagle`s MEM (Eagle`s Minimum Essential Medium)DMEM( Dulbcco's MEM )-- exMCDB *

Phenol red*

- 1.a. Na+K+ Cl- b. Mg2+ ()c. Ca2+ d. H2PO4- pH H+ HPO42- H2PO4- OH- *

- 2. : L-Histidine-HCl-H2O L-Isoleucine L-Leucine L-Lysine-HCl L-Methionine L-Phenylalanine L-Threonine L-Tryptophan L-Valine : L-Alanine L-Arginine-HCl L-Asparagine-H2O L-Aspartic Acid L-Cystine-2HCl L-Glutamic Acid L-Glutamine Glycine L-Proline L-Serine L-Tyrosine-2Na-2H2O*

- 3. ,,*

- 4. RNADNA

pH6.6-7.8 pH7.2-7.4: - 5. *

- 6.*

Penicillin100 unit/ml-20G(+)bacteriaStreptomycin100 g/ml -20G(+-)bacteriaChlotetracycline50 g/ml-20G(+-)bacteriaGentamycin50g/ml-20G(+-)bacteriaMycoplasmaAmphotericicin B2.5g/ml-20YeastmoldsNystatin50g/ml-20Yeastmoldsfungizone2.5g/ml-20Yeastmolds

a.(growth factor) (Epidermal Growth FactorEGF) b.(hormones) , c.(attachment factor) , fibronectin - 7. *

d. (binding protein:albumin) , Transferrine. (lipid source) Phospholipidsf. (trace element) Cu, Zn, Co, Mo, Se,coenzyme,Zn

* - 7.

(carbon sources)(nitrogen sources)http://ezphysics.nchu.edu.tw/genphys/gen/92to95/handout/1.pdf *

(Natural media)(Basic media)(Selective media)EMB agarE.coli(Differential media)http://microbiology.scu.edu.tw/micro/microbe-exp/exp_website/Media.htm

*

(enriched media)(Synthetic media)chemically defined medium()

*

(Complex media)(Yeast extract)(Peptone) (1)(Nutrient brothNB)(Beef extract)(Peptone)(Nutrient agarNA) (2)(Brian Heart InfusionBHI)(Infusion)(Fastidious) (3)(Lysogeny broth LB) (Yeast extract)(Tryptone)*

*

(Broth)(Agar)(Plate) (Slant)(agar deep tube)0.5~0.8%90 10042 *

pH*

*

*

*

pH6.5~7.559

(halophiles)(saccharophiles) (osmophiles)http://www1.tf.edu.tw/top/department/food/%E9%A3%9F%E5%93%81%E7%B3%BB%E7%B6%B2%E9%A0%81/teacher&research/%E9%99%B3%E5%A7%BF%E5%88%A9%E8%80%81%E5%B8%AB/lesson01/03%E5%BE%AE%E7%94%9F%E7%89%A9%E7%9A%84%E5%9F%B9%E9%A4%8A.pdf*

University of Massachusetts at AmherstSwades ChaudhuriDerek R. Lovley Rhodoferax ferrireducens80

http://sa.ylib.com/news/newsshow.asp?FDocNo=358&CL=26*

DEP

Low conductivity Sucrose D-mannitol( =5 S/cm) ( =1 S/cm) HEPES( =60 S/cm)

High conductivity PBS( =16000 S/cm)*

*

molecular genetics/

*

http://www.supermt.com.tw/molbio.htm*

Thank you for your attention*

********acetic acid *Boric acid **citric acid *pH*facultative anaerobes NO3SO42 aerotolerant anaerobes superoxide dismutasecatalase *