Line 100: | Line 100: | ||
<div class="container"> | <div class="container"> | ||
<div class="section_title "> | <div class="section_title "> | ||
− | + | <h1 align="center" >Protocols </h1><hr> | |
− | <table border="1" align="center" cellspacing="0" > | + | <div align="center"><table border="1" align="center" cellspacing="0" > |
<tr> | <tr> | ||
<td width="115" valign="top" ><br> | <td width="115" valign="top" ><br> | ||
Line 127: | Line 127: | ||
<td width="710" valign="top" colspan="2" ><p style="font-family: 'Arial Unicode MS', 'Microsoft YaHei UI', 'Microsoft YaHei UI Light', '华文细黑', '微软雅黑', '幼圆'; font-size: 18px;"> Adjustable ph: 1l plus 200μl,5mol L-1NaOH ph to 7.0 </p></td> | <td width="710" valign="top" colspan="2" ><p style="font-family: 'Arial Unicode MS', 'Microsoft YaHei UI', 'Microsoft YaHei UI Light', '华文细黑', '微软雅黑', '幼圆'; font-size: 18px;"> Adjustable ph: 1l plus 200μl,5mol L-1NaOH ph to 7.0 </p></td> | ||
</tr> | </tr> | ||
− | </ | + | </div> |
<h2 style="text-align: center" >LB Srednje(1L solid) </h2> | <h2 style="text-align: center" >LB Srednje(1L solid) </h2> | ||
− | <table border="1" align="center" cellspacing="0" > | + | <div align="center"><table border="1" align="center" cellspacing="0" > |
<tr> | <tr> | ||
<td width="115" valign="top" ><p style="font-family: 'Arial Unicode MS', 'Microsoft YaHei UI', 'Microsoft YaHei UI Light', '华文细黑', '微软雅黑', '幼圆'; font-size: 18px;"> | <td width="115" valign="top" ><p style="font-family: 'Arial Unicode MS', 'Microsoft YaHei UI', 'Microsoft YaHei UI Light', '华文细黑', '微软雅黑', '幼圆'; font-size: 18px;"> | ||
Line 154: | Line 154: | ||
<td width="710" valign="top" colspan="2" ><p style="font-family: 'Arial Unicode MS', 'Microsoft YaHei UI', 'Microsoft YaHei UI Light', '华文细黑', '微软雅黑', '幼圆'; font-size: 18px;"> Adjustable ph: 1l plus 200μl,5mol L-1 NaOH ph to 7.0 </p></td> | <td width="710" valign="top" colspan="2" ><p style="font-family: 'Arial Unicode MS', 'Microsoft YaHei UI', 'Microsoft YaHei UI Light', '华文细黑', '微软雅黑', '幼圆'; font-size: 18px;"> Adjustable ph: 1l plus 200μl,5mol L-1 NaOH ph to 7.0 </p></td> | ||
</tr> | </tr> | ||
− | </ | + | </div> |
<p style="font-family: 'Arial Unicode MS', 'Microsoft YaHei UI', 'Microsoft YaHei UI Light', '华文细黑', '微软雅黑', '幼圆'; font-size: 18px;"> </p> | <p style="font-family: 'Arial Unicode MS', 'Microsoft YaHei UI', 'Microsoft YaHei UI Light', '华文细黑', '微软雅黑', '幼圆'; font-size: 18px;"> </p> | ||
− | <table border="1" align="center" cellspacing="0" > | + | <div align="center"><table border="1" align="center" cellspacing="0" > |
<tr> | <tr> | ||
<td width="174" valign="top" ><p style="font-family: 'Arial Unicode MS', 'Microsoft YaHei UI', 'Microsoft YaHei UI Light', '华文细黑', '微软雅黑', '幼圆'; font-size: 18px;"> 50*TAE(1L) </p></td> | <td width="174" valign="top" ><p style="font-family: 'Arial Unicode MS', 'Microsoft YaHei UI', 'Microsoft YaHei UI Light', '华文细黑', '微软雅黑', '幼圆'; font-size: 18px;"> 50*TAE(1L) </p></td> | ||
Line 181: | Line 181: | ||
<td width="535" valign="top" ><p style="font-family: 'Arial Unicode MS', 'Microsoft YaHei UI', 'Microsoft YaHei UI Light', '华文细黑', '微软雅黑', '幼圆'; font-size: 18px;"> </p></td> | <td width="535" valign="top" ><p style="font-family: 'Arial Unicode MS', 'Microsoft YaHei UI', 'Microsoft YaHei UI Light', '华文细黑', '微软雅黑', '幼圆'; font-size: 18px;"> </p></td> | ||
</tr> | </tr> | ||
− | </ | + | </div> |
<p style="font-family: 'Arial Unicode MS', 'Microsoft YaHei UI', 'Microsoft YaHei UI Light', '华文细黑', '微软雅黑', '幼圆'; font-size: 18px;"> </p> | <p style="font-family: 'Arial Unicode MS', 'Microsoft YaHei UI', 'Microsoft YaHei UI Light', '华文细黑', '微软雅黑', '幼圆'; font-size: 18px;"> </p> | ||
− | <table border="1" align="center" cellspacing="0" > | + | <div align="center"><table border="1" align="center" cellspacing="0" > |
<tr> | <tr> | ||
<td width="162" valign="top" ><p style="font-family: 'Arial Unicode MS', 'Microsoft YaHei UI', 'Microsoft YaHei UI Light', '华文细黑', '微软雅黑', '幼圆'; font-size: 18px;"> 0.25 % dye(1L) </p></td> | <td width="162" valign="top" ><p style="font-family: 'Arial Unicode MS', 'Microsoft YaHei UI', 'Microsoft YaHei UI Light', '华文细黑', '微软雅黑', '幼圆'; font-size: 18px;"> 0.25 % dye(1L) </p></td> | ||
Line 204: | Line 204: | ||
<td width="547" valign="top" ><p style="font-family: 'Arial Unicode MS', 'Microsoft YaHei UI', 'Microsoft YaHei UI Light', '华文细黑', '微软雅黑', '幼圆'; font-size: 18px;"> </p></td> | <td width="547" valign="top" ><p style="font-family: 'Arial Unicode MS', 'Microsoft YaHei UI', 'Microsoft YaHei UI Light', '华文细黑', '微软雅黑', '幼圆'; font-size: 18px;"> </p></td> | ||
</tr> | </tr> | ||
− | </ | + | </div> |
<p style="font-family: 'Arial Unicode MS', 'Microsoft YaHei UI', 'Microsoft YaHei UI Light', '华文细黑', '微软雅黑', '幼圆'; font-size: 18px;"> </p> | <p style="font-family: 'Arial Unicode MS', 'Microsoft YaHei UI', 'Microsoft YaHei UI Light', '华文细黑', '微软雅黑', '幼圆'; font-size: 18px;"> </p> | ||
− | <table border="1" align="center" cellspacing="0" > | + | <div align="center"><table border="1" align="center" cellspacing="0" > |
<tr > | <tr > | ||
<td width="174" valign="top" ><p style="font-family: 'Arial Unicode MS', 'Microsoft YaHei UI', 'Microsoft YaHei UI Light', '华文细黑', '微软雅黑', '幼圆'; font-size: 18px;"> 10 % seperate adhesive(10ml) </p></td> | <td width="174" valign="top" ><p style="font-family: 'Arial Unicode MS', 'Microsoft YaHei UI', 'Microsoft YaHei UI Light', '华文细黑', '微软雅黑', '幼圆'; font-size: 18px;"> 10 % seperate adhesive(10ml) </p></td> | ||
Line 235: | Line 235: | ||
<td width="534" valign="top" ><p style="font-family: 'Arial Unicode MS', 'Microsoft YaHei UI', 'Microsoft YaHei UI Light', '华文细黑', '微软雅黑', '幼圆'; font-size: 18px;"> 0.002ml </p></td> | <td width="534" valign="top" ><p style="font-family: 'Arial Unicode MS', 'Microsoft YaHei UI', 'Microsoft YaHei UI Light', '华文细黑', '微软雅黑', '幼圆'; font-size: 18px;"> 0.002ml </p></td> | ||
</tr> | </tr> | ||
− | </ | + | </div> |
<p style="font-family: 'Arial Unicode MS', 'Microsoft YaHei UI', 'Microsoft YaHei UI Light', '华文细黑', '微软雅黑', '幼圆'; font-size: 18px;"> </p> | <p style="font-family: 'Arial Unicode MS', 'Microsoft YaHei UI', 'Microsoft YaHei UI Light', '华文细黑', '微软雅黑', '幼圆'; font-size: 18px;"> </p> | ||
− | <table border="1" align="center" cellspacing="0" > | + | <div align="center"><table border="1" align="center" cellspacing="0" > |
<tr > | <tr > | ||
<td width="174" valign="top" ><p style="font-family: 'Arial Unicode MS', 'Microsoft YaHei UI', 'Microsoft YaHei UI Light', '华文细黑', '微软雅黑', '幼圆'; font-size: 18px;"> 5 % concentrate glue(6ml) </p></td> | <td width="174" valign="top" ><p style="font-family: 'Arial Unicode MS', 'Microsoft YaHei UI', 'Microsoft YaHei UI Light', '华文细黑', '微软雅黑', '幼圆'; font-size: 18px;"> 5 % concentrate glue(6ml) </p></td> | ||
Line 266: | Line 266: | ||
<td width="534" valign="top" ><p style="font-family: 'Arial Unicode MS', 'Microsoft YaHei UI', 'Microsoft YaHei UI Light', '华文细黑', '微软雅黑', '幼圆'; font-size: 18px;"> 0.004ml </p></td> | <td width="534" valign="top" ><p style="font-family: 'Arial Unicode MS', 'Microsoft YaHei UI', 'Microsoft YaHei UI Light', '华文细黑', '微软雅黑', '幼圆'; font-size: 18px;"> 0.004ml </p></td> | ||
</tr> | </tr> | ||
− | </ | + | </div> |
<p style="font-family: 'Arial Unicode MS', 'Microsoft YaHei UI', 'Microsoft YaHei UI Light', '华文细黑', '微软雅黑', '幼圆'; font-size: 18px;"> </p> | <p style="font-family: 'Arial Unicode MS', 'Microsoft YaHei UI', 'Microsoft YaHei UI Light', '华文细黑', '微软雅黑', '幼圆'; font-size: 18px;"> </p> | ||
<p style="font-family: 'Arial Unicode MS', 'Microsoft YaHei UI', 'Microsoft YaHei UI Light', '华文细黑', '微软雅黑', '幼圆'; font-size: 18px; text-align: center;"> Nickel column purification buffer <br> | <p style="font-family: 'Arial Unicode MS', 'Microsoft YaHei UI', 'Microsoft YaHei UI Light', '华文细黑', '微软雅黑', '幼圆'; font-size: 18px; text-align: center;"> Nickel column purification buffer <br> | ||
Line 287: | Line 287: | ||
<span style="text-align: center"> Enzyme buffer solution: | <span style="text-align: center"> Enzyme buffer solution: | ||
</h2> | </h2> | ||
− | <table border="1" align="center" cellspacing="0" > | + | <div align="center"><table border="1" align="center" cellspacing="0" > |
<tr > | <tr > | ||
<td width="133" valign="top" ><br> | <td width="133" valign="top" ><br> | ||
Line 313: | Line 313: | ||
<td width="382" valign="top" ><p style="font-family: 'Arial Unicode MS', 'Microsoft YaHei UI', 'Microsoft YaHei UI Light', '华文细黑', '微软雅黑', '幼圆'; font-size: 18px;"> 20μL </p></td> | <td width="382" valign="top" ><p style="font-family: 'Arial Unicode MS', 'Microsoft YaHei UI', 'Microsoft YaHei UI Light', '华文细黑', '微软雅黑', '幼圆'; font-size: 18px;"> 20μL </p></td> | ||
</tr> | </tr> | ||
− | </ | + | </div> |
<h2 style="text-align: center" >P-nitrophenol with different concentrations </h2> | <h2 style="text-align: center" >P-nitrophenol with different concentrations </h2> | ||
− | <table border="1" align="center" cellspacing="0" > | + | <div align="center"><table border="1" align="center" cellspacing="0" > |
<tr> | <tr> | ||
<td width="103" valign="top" ><br> | <td width="103" valign="top" ><br> | ||
Line 341: | Line 341: | ||
<td width="275" valign="top" ><p style="font-family: 'Arial Unicode MS', 'Microsoft YaHei UI', 'Microsoft YaHei UI Light', '华文细黑', '微软雅黑', '幼圆'; font-size: 18px;"> 10 mL </p></td> | <td width="275" valign="top" ><p style="font-family: 'Arial Unicode MS', 'Microsoft YaHei UI', 'Microsoft YaHei UI Light', '华文细黑', '微软雅黑', '幼圆'; font-size: 18px;"> 10 mL </p></td> | ||
</tr> | </tr> | ||
− | </ | + | </div> |
<p style="font-family: 'Arial Unicode MS', 'Microsoft YaHei UI', 'Microsoft YaHei UI Light', '华文细黑', '微软雅黑', '幼圆'; font-size: 18px;"> </p> | <p style="font-family: 'Arial Unicode MS', 'Microsoft YaHei UI', 'Microsoft YaHei UI Light', '华文细黑', '微软雅黑', '幼圆'; font-size: 18px;"> </p> | ||
<h2> DNA gum recovery:</h2> | <h2> DNA gum recovery:</h2> | ||
Line 497: | Line 497: | ||
7) after 3 min, the absorbance value is measured by ultraviolet spectrophotometer at the wavelength of three nm; <br> | 7) after 3 min, the absorbance value is measured by ultraviolet spectrophotometer at the wavelength of three nm; <br> | ||
8) Compare the standard curve and calculate the conversion Rate. Enzyme activity is defined as: at room temperature, hydrolysis of 1μmol acetate per minute of the amount of enzyme required for nitrobenzene ester is an enzyme unit. </p> | 8) Compare the standard curve and calculate the conversion Rate. Enzyme activity is defined as: at room temperature, hydrolysis of 1μmol acetate per minute of the amount of enzyme required for nitrobenzene ester is an enzyme unit. </p> | ||
+ | |||
Revision as of 15:43, 14 October 2018
![](https://static.igem.org/mediawiki/2018/c/c1/T--AHUT_China--_chilun.jpg)
Protocols
LB Medium(1L): |
|
||||
tryptone |
10g |
||||
yeast extract |
5g |
||||
NaCl |
10g |
||||
ddH2O |
|
||||
Adjustable ph: 1l plus 200μl,5mol L-1NaOH ph to 7.0 |
tryptone |
10g |
||||
yeast extract |
5g |
||||
NaCl |
10g |
||||
agar |
15g |
||||
ddH2O |
|
||||
Adjustable ph: 1l plus 200μl,5mol L-1 NaOH ph to 7.0 |
50*TAE(1L) |
|
||||
ddH2O |
500ml |
||||
Tris buffer solution |
242g |
||||
0.5mol·l-1 EDTA |
100ml |
||||
Glacial acetic acid |
57.1ml |
||||
ddH2O |
|
0.25 % dye(1L) |
|
||||
G-250 |
100mg |
||||
90%ethanol |
50ml |
||||
85% phosphoric acid |
10ml |
||||
ddH2O |
|
10 % seperate adhesive(10ml) |
|
||||
distilled water |
1.9ml |
||||
30%Arc-Bis |
1.7ml |
||||
1.5M Tris-HCl pH 8.8 |
1.3ml |
||||
10%SDS |
0.05ml |
||||
10 % ammonium persulfate |
0.05ml |
||||
TEMED |
0.002ml |
5 % concentrate glue(6ml) |
|
||||
distilled water |
2.7ml |
||||
30%Arc-Bis |
0.67ml |
||||
1.0M Tris-HCl pH 6.8 |
0.5ml |
||||
10%SDS |
0.04ml |
||||
10 % ammonium persulfate |
0.04ml |
||||
TEMED |
0.004ml |
Buffer solution |
1ml |
990 μL |
980 μL |
970 μL |
960 μL |
Enzyme(1 μg/mL) |
0μL |
10μL |
20μL |
30μL |
40μL |
Bromothyme blue indicator |
20μL |
20μL |
20μL |
20μL |
20μL |
concentration |
0.01 mM |
0.02 mM |
0.03 mM |
0.04 mM |
0.05 mM |
p-NP(0.1 mM) |
2 mL |
4 mL |
6 mL |
8 mL |
10 mL |
water |
18 mL |
16 mL |
14 mL |
12 mL |
10 mL |