IGEM:Hong Kong UST 2014/2009/Notebook/ATP Concentration Effect to Ligation Speed/2014/06/05: Difference between revisions
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== | ==Thursday, 5th of June 2014== | ||
* | * '''Making the ATP Stock'''<br> | ||
Make 10 mM ATP stock for 1 ml by doing serial dilution.<br> | |||
1. Weigh out 0.06 grams of powdered ATP and dilute it using 1 ml of water.<br> | |||
2. Pipette 100 µl of the diluted ATP powder and transfer it into a micro centrifuge tube.<br> | |||
3. Add 900 µl of double distilled water to make the 10mM ATP stock.<br><br> | |||
*'''Ligation'''<br> | |||
Make the recipe for ligation. Each person in the team will use 4 micro centrifuge tube to create the control, 1mM ATP, 2mM ATP, and 4 mM ATP. The amount of insert and backbone added varies due to the different amount of concentration of nucleic acid in each set. <br> | |||
The ratio of insert to backbone must be 3:1. And since the length of the insert and backbone is 1 kb to 2 kb respectively, the amount of insert must be 30 ng and backbone must be 20 ng. ( total amount of both insert and backbone is ~50 ng)<br><br> | |||
<span id="docs-internal-guid-f1862da3-6fcc-ab8c-ee21-a7d5a17346e7"><br /></span> | |||
<br /> | |||
<div dir="ltr" style="margin-left: 0pt;"> | |||
<table style="border-collapse: collapse; border: none;"> | |||
<tr style="height: 0px;"><td style="border-bottom: solid #000000 1px; border-left: solid #000000 1px; border-right: solid #000000 1px; border-top: solid #000000 1px; padding: 7px 7px 7px 7px; vertical-align: top;"><span style="font-size: x-small;"><br /></span></td><td style="border-bottom: solid #000000 1px; border-left: solid #000000 1px; border-right: solid #000000 1px; border-top: solid #000000 1px; padding: 7px 7px 7px 7px; vertical-align: top;"><div dir="ltr" style="line-height: 1.15; margin-bottom: 0pt; margin-top: 0pt; text-align: center;"> | |||
<span style="background-color: transparent; color: black; font-family: Arial; font-size: x-small; font-style: normal; font-variant: normal; font-weight: bold; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">Control</span></div> | |||
</td><td style="border-bottom: solid #000000 1px; border-left: solid #000000 1px; border-right: solid #000000 1px; border-top: solid #000000 1px; padding: 7px 7px 7px 7px; vertical-align: top;"><div dir="ltr" style="line-height: 1.15; margin-bottom: 0pt; margin-top: 0pt; text-align: center;"> | |||
<span style="background-color: transparent; color: black; font-family: Arial; font-size: x-small; font-style: normal; font-variant: normal; font-weight: bold; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">1mM ATP</span></div> | |||
</td><td style="border-bottom: solid #000000 1px; border-left: solid #000000 1px; border-right: solid #000000 1px; border-top: solid #000000 1px; padding: 7px 7px 7px 7px; vertical-align: top;"><div dir="ltr" style="line-height: 1.15; margin-bottom: 0pt; margin-top: 0pt; text-align: center;"> | |||
<span style="background-color: transparent; color: black; font-family: Arial; font-size: x-small; font-style: normal; font-variant: normal; font-weight: bold; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">2mM ATP</span></div> | |||
</td><td style="border-bottom: solid #000000 1px; border-left: solid #000000 1px; border-right: solid #000000 1px; border-top: solid #000000 1px; padding: 7px 7px 7px 7px; vertical-align: top;"><div dir="ltr" style="line-height: 1.15; margin-bottom: 0pt; margin-top: 0pt; text-align: center;"> | |||
<span style="background-color: transparent; color: black; font-family: Arial; font-size: x-small; font-style: normal; font-variant: normal; font-weight: bold; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">4mM ATP</span></div> | |||
</td></tr> | |||
<tr style="height: 0px;"><td style="border-bottom: solid #000000 1px; border-left: solid #000000 1px; border-right: solid #000000 1px; border-top: solid #000000 1px; padding: 7px 7px 7px 7px; vertical-align: top;"><div dir="ltr" style="line-height: 1.15; margin-bottom: 0pt; margin-top: 0pt; text-align: center;"> | |||
<span style="background-color: transparent; color: black; font-family: Arial; font-size: x-small; font-style: normal; font-variant: normal; font-weight: bold; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">Insert</span></div> | |||
</td><td style="border-bottom: solid #000000 1px; border-left: solid #000000 1px; border-right: solid #000000 1px; border-top: solid #000000 1px; padding: 7px 7px 7px 7px; vertical-align: top;"><div dir="ltr" style="line-height: 1.15; margin-bottom: 0pt; margin-top: 0pt; text-align: center;"> | |||
<span style="background-color: transparent; color: black; font-family: Arial; font-size: x-small; font-style: normal; font-variant: normal; font-weight: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">30.0 ng</span></div> | |||
</td><td style="border-bottom: solid #000000 1px; border-left: solid #000000 1px; border-right: solid #000000 1px; border-top: solid #000000 1px; padding: 7px 7px 7px 7px; vertical-align: top;"><div dir="ltr" style="line-height: 1.15; margin-bottom: 0pt; margin-top: 0pt; text-align: center;"> | |||
<span style="background-color: transparent; color: black; font-family: Arial; font-size: x-small; font-style: normal; font-variant: normal; font-weight: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">30.0 ng</span></div> | |||
</td><td style="border-bottom: solid #000000 1px; border-left: solid #000000 1px; border-right: solid #000000 1px; border-top: solid #000000 1px; padding: 7px 7px 7px 7px; vertical-align: top;"><div dir="ltr" style="line-height: 1.15; margin-bottom: 0pt; margin-top: 0pt; text-align: center;"> | |||
<span style="background-color: transparent; color: black; font-family: Arial; font-size: x-small; font-style: normal; font-variant: normal; font-weight: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">30.0 ng</span></div> | |||
</td><td style="border-bottom: solid #000000 1px; border-left: solid #000000 1px; border-right: solid #000000 1px; border-top: solid #000000 1px; padding: 7px 7px 7px 7px; vertical-align: top;"><div dir="ltr" style="line-height: 1.15; margin-bottom: 0pt; margin-top: 0pt; text-align: center;"> | |||
<span style="background-color: transparent; color: black; font-family: Arial; font-size: x-small; font-style: normal; font-variant: normal; font-weight: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">30.0 ng</span></div> | |||
</td></tr> | |||
<tr style="height: 0px;"><td style="border-bottom: solid #000000 1px; border-left: solid #000000 1px; border-right: solid #000000 1px; border-top: solid #000000 1px; padding: 7px 7px 7px 7px; vertical-align: top;"><div dir="ltr" style="line-height: 1.15; margin-bottom: 0pt; margin-top: 0pt; text-align: center;"> | |||
<span style="background-color: transparent; color: black; font-family: Arial; font-size: x-small; font-style: normal; font-variant: normal; font-weight: bold; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">Backbone</span></div> | |||
</td><td style="border-bottom: solid #000000 1px; border-left: solid #000000 1px; border-right: solid #000000 1px; border-top: solid #000000 1px; padding: 7px 7px 7px 7px; vertical-align: top;"><div dir="ltr" style="line-height: 1.15; margin-bottom: 0pt; margin-top: 0pt; text-align: center;"> | |||
<span style="background-color: transparent; color: black; font-family: Arial; font-size: x-small; font-style: normal; font-variant: normal; font-weight: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">20.0 ng</span></div> | |||
</td><td style="border-bottom: solid #000000 1px; border-left: solid #000000 1px; border-right: solid #000000 1px; border-top: solid #000000 1px; padding: 7px 7px 7px 7px; vertical-align: top;"><div dir="ltr" style="line-height: 1.15; margin-bottom: 0pt; margin-top: 0pt; text-align: center;"> | |||
<span style="background-color: transparent; color: black; font-family: Arial; font-size: x-small; font-style: normal; font-variant: normal; font-weight: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">20.0 ng</span></div> | |||
</td><td style="border-bottom: solid #000000 1px; border-left: solid #000000 1px; border-right: solid #000000 1px; border-top: solid #000000 1px; padding: 7px 7px 7px 7px; vertical-align: top;"><div dir="ltr" style="line-height: 1.15; margin-bottom: 0pt; margin-top: 0pt; text-align: center;"> | |||
<span style="background-color: transparent; color: black; font-family: Arial; font-size: x-small; font-style: normal; font-variant: normal; font-weight: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">20.0 ng</span></div> | |||
</td><td style="border-bottom: solid #000000 1px; border-left: solid #000000 1px; border-right: solid #000000 1px; border-top: solid #000000 1px; padding: 7px 7px 7px 7px; vertical-align: top;"><div dir="ltr" style="line-height: 1.15; margin-bottom: 0pt; margin-top: 0pt; text-align: center;"> | |||
<span style="background-color: transparent; color: black; font-family: Arial; font-size: x-small; font-style: normal; font-variant: normal; font-weight: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">20.0 ng</span></div> | |||
</td></tr> | |||
<tr style="height: 0px;"><td style="border-bottom: solid #000000 1px; border-left: solid #000000 1px; border-right: solid #000000 1px; border-top: solid #000000 1px; padding: 7px 7px 7px 7px; vertical-align: top;"><div dir="ltr" style="line-height: 1.15; margin-bottom: 0pt; margin-top: 0pt; text-align: center;"> | |||
<span style="font-size: x-small;"><span style="background-color: transparent; color: black; font-family: Arial; font-style: normal; font-variant: normal; font-weight: bold; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">10x T4 buffer (contain 1mM of ATP) (</span><span style="background-color: white; color: black; font-family: Arial; font-style: normal; font-variant: normal; font-weight: bold; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">µl</span><span style="background-color: transparent; color: black; font-family: Arial; font-style: normal; font-variant: normal; font-weight: bold; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">)</span></span></div> | |||
</td><td style="border-bottom: solid #000000 1px; border-left: solid #000000 1px; border-right: solid #000000 1px; border-top: solid #000000 1px; padding: 7px 7px 7px 7px; vertical-align: top;"><div dir="ltr" style="line-height: 1.15; margin-bottom: 0pt; margin-top: 0pt; text-align: center;"> | |||
<span style="background-color: transparent; color: black; font-family: Arial; font-size: x-small; font-style: normal; font-variant: normal; font-weight: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">-</span></div> | |||
</td><td style="border-bottom: solid #000000 1px; border-left: solid #000000 1px; border-right: solid #000000 1px; border-top: solid #000000 1px; padding: 7px 7px 7px 7px; vertical-align: top;"><div dir="ltr" style="line-height: 1.15; margin-bottom: 0pt; margin-top: 0pt; text-align: center;"> | |||
<span style="background-color: transparent; color: black; font-family: Arial; font-size: x-small; font-style: normal; font-variant: normal; font-weight: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">5.0</span></div> | |||
</td><td style="border-bottom: solid #000000 1px; border-left: solid #000000 1px; border-right: solid #000000 1px; border-top: solid #000000 1px; padding: 7px 7px 7px 7px; vertical-align: top;"><div dir="ltr" style="line-height: 1.15; margin-bottom: 0pt; margin-top: 0pt; text-align: center;"> | |||
<span style="background-color: transparent; color: black; font-family: Arial; font-size: x-small; font-style: normal; font-variant: normal; font-weight: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">5.0</span></div> | |||
</td><td style="border-bottom: solid #000000 1px; border-left: solid #000000 1px; border-right: solid #000000 1px; border-top: solid #000000 1px; padding: 7px 7px 7px 7px; vertical-align: top;"><div dir="ltr" style="line-height: 1.15; margin-bottom: 0pt; margin-top: 0pt; text-align: center;"> | |||
<span style="background-color: transparent; color: black; font-family: Arial; font-size: x-small; font-style: normal; font-variant: normal; font-weight: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">5.0</span></div> | |||
</td></tr> | |||
<tr style="height: 0px;"><td style="border-bottom: solid #000000 1px; border-left: solid #000000 1px; border-right: solid #000000 1px; border-top: solid #000000 1px; padding: 7px 7px 7px 7px; vertical-align: top;"><div dir="ltr" style="line-height: 1.15; margin-bottom: 0pt; margin-top: 0pt; text-align: center;"> | |||
<span style="font-size: x-small;"><span style="background-color: transparent; color: black; font-family: Arial; font-style: normal; font-variant: normal; font-weight: bold; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">10 mM ATP (</span><span style="background-color: white; color: black; font-family: Arial; font-style: normal; font-variant: normal; font-weight: bold; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">µl</span><span style="background-color: transparent; color: black; font-family: Arial; font-style: normal; font-variant: normal; font-weight: bold; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">)</span></span></div> | |||
</td><td style="border-bottom: solid #000000 1px; border-left: solid #000000 1px; border-right: solid #000000 1px; border-top: solid #000000 1px; padding: 7px 7px 7px 7px; vertical-align: top;"><div dir="ltr" style="line-height: 1.15; margin-bottom: 0pt; margin-top: 0pt; text-align: center;"> | |||
<span style="background-color: transparent; color: black; font-family: Arial; font-size: x-small; font-style: normal; font-variant: normal; font-weight: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">-</span></div> | |||
</td><td style="border-bottom: solid #000000 1px; border-left: solid #000000 1px; border-right: solid #000000 1px; border-top: solid #000000 1px; padding: 7px 7px 7px 7px; vertical-align: top;"><div dir="ltr" style="line-height: 1.15; margin-bottom: 0pt; margin-top: 0pt; text-align: center;"> | |||
<span style="background-color: transparent; color: black; font-family: Arial; font-size: x-small; font-style: normal; font-variant: normal; font-weight: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">-</span></div> | |||
</td><td style="border-bottom: solid #000000 1px; border-left: solid #000000 1px; border-right: solid #000000 1px; border-top: solid #000000 1px; padding: 7px 7px 7px 7px; vertical-align: top;"><div dir="ltr" style="line-height: 1.15; margin-bottom: 0pt; margin-top: 0pt; text-align: center;"> | |||
<span style="background-color: transparent; color: black; font-family: Arial; font-size: x-small; font-style: normal; font-variant: normal; font-weight: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">5.0</span></div> | |||
</td><td style="border-bottom: solid #000000 1px; border-left: solid #000000 1px; border-right: solid #000000 1px; border-top: solid #000000 1px; padding: 7px 7px 7px 7px; vertical-align: top;"><div dir="ltr" style="line-height: 1.15; margin-bottom: 0pt; margin-top: 0pt; text-align: center;"> | |||
<span style="background-color: transparent; color: black; font-family: Arial; font-size: x-small; font-style: normal; font-variant: normal; font-weight: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">15.0</span></div> | |||
</td></tr> | |||
<tr style="height: 0px;"><td style="border-bottom: solid #000000 1px; border-left: solid #000000 1px; border-right: solid #000000 1px; border-top: solid #000000 1px; padding: 7px 7px 7px 7px; vertical-align: top;"><div dir="ltr" style="line-height: 1.15; margin-bottom: 0pt; margin-top: 0pt; text-align: center;"> | |||
<span style="font-size: x-small;"><span style="background-color: transparent; color: black; font-family: Arial; font-style: normal; font-variant: normal; font-weight: bold; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">T4 Ligase (</span><span style="background-color: white; color: black; font-family: Arial; font-style: normal; font-variant: normal; font-weight: bold; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">µl</span><span style="background-color: transparent; color: black; font-family: Arial; font-style: normal; font-variant: normal; font-weight: bold; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">)</span></span></div> | |||
</td><td style="border-bottom: solid #000000 1px; border-left: solid #000000 1px; border-right: solid #000000 1px; border-top: solid #000000 1px; padding: 7px 7px 7px 7px; vertical-align: top;"><div dir="ltr" style="line-height: 1.15; margin-bottom: 0pt; margin-top: 0pt; text-align: center;"> | |||
<span style="background-color: transparent; color: black; font-family: Arial; font-size: x-small; font-style: normal; font-variant: normal; font-weight: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">0.5</span></div> | |||
</td><td style="border-bottom: solid #000000 1px; border-left: solid #000000 1px; border-right: solid #000000 1px; border-top: solid #000000 1px; padding: 7px 7px 7px 7px; vertical-align: top;"><div dir="ltr" style="line-height: 1.15; margin-bottom: 0pt; margin-top: 0pt; text-align: center;"> | |||
<span style="background-color: transparent; color: black; font-family: Arial; font-size: x-small; font-style: normal; font-variant: normal; font-weight: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">0.5</span></div> | |||
</td><td style="border-bottom: solid #000000 1px; border-left: solid #000000 1px; border-right: solid #000000 1px; border-top: solid #000000 1px; padding: 7px 7px 7px 7px; vertical-align: top;"><div dir="ltr" style="line-height: 1.15; margin-bottom: 0pt; margin-top: 0pt; text-align: center;"> | |||
<span style="background-color: transparent; color: black; font-family: Arial; font-size: x-small; font-style: normal; font-variant: normal; font-weight: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">0.5</span></div> | |||
</td><td style="border-bottom: solid #000000 1px; border-left: solid #000000 1px; border-right: solid #000000 1px; border-top: solid #000000 1px; padding: 7px 7px 7px 7px; vertical-align: top;"><div dir="ltr" style="line-height: 1.15; margin-bottom: 0pt; margin-top: 0pt; text-align: center;"> | |||
<span style="background-color: transparent; color: black; font-family: Arial; font-size: x-small; font-style: normal; font-variant: normal; font-weight: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">0.5</span></div> | |||
</td></tr> | |||
<tr style="height: 0px;"><td style="border-bottom: solid #000000 1px; border-left: solid #000000 1px; border-right: solid #000000 1px; border-top: solid #000000 1px; padding: 7px 7px 7px 7px; vertical-align: top;"><div dir="ltr" style="margin-bottom: 0pt; margin-top: 0pt; text-align: center;"> | |||
<span style="font-family: Arial; font-size: x-small;"><span style="line-height: 14.949999809265137px; white-space: pre-wrap;"><b>ddH2O</b></span></span></div> | |||
</td><td style="border-bottom: solid #000000 1px; border-left: solid #000000 1px; border-right: solid #000000 1px; border-top: solid #000000 1px; padding: 7px 7px 7px 7px; vertical-align: top;"><div dir="ltr" style="line-height: 1.15; margin-bottom: 0pt; margin-top: 0pt; text-align: center;"> | |||
<span style="background-color: transparent; color: black; font-family: Arial; font-size: x-small; font-style: italic; font-variant: normal; font-weight: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">Adjust to total volume</span></div> | |||
</td><td style="border-bottom: solid #000000 1px; border-left: solid #000000 1px; border-right: solid #000000 1px; border-top: solid #000000 1px; padding: 7px 7px 7px 7px; vertical-align: top;"><div dir="ltr" style="line-height: 1.15; margin-bottom: 0pt; margin-top: 0pt; text-align: center;"> | |||
<span style="background-color: transparent; color: black; font-family: Arial; font-size: x-small; font-style: italic; font-variant: normal; font-weight: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">Adjust to total volume</span></div> | |||
</td><td style="border-bottom: solid #000000 1px; border-left: solid #000000 1px; border-right: solid #000000 1px; border-top: solid #000000 1px; padding: 7px 7px 7px 7px; vertical-align: top;"><div dir="ltr" style="line-height: 1.15; margin-bottom: 0pt; margin-top: 0pt; text-align: center;"> | |||
<span style="font-family: Arial; font-size: x-small; font-style: italic; line-height: 14.949999809265137px; white-space: pre-wrap;">Adjust to total volume</span></div> | |||
</td><td style="border-bottom: solid #000000 1px; border-left: solid #000000 1px; border-right: solid #000000 1px; border-top: solid #000000 1px; padding: 7px 7px 7px 7px; vertical-align: top;"><div dir="ltr" style="line-height: 1.15; margin-bottom: 0pt; margin-top: 0pt; text-align: center;"> | |||
<span style="font-family: Arial; font-size: x-small; font-style: italic; line-height: 14.949999809265137px; white-space: pre-wrap;">Adjust to total volume</span></div> | |||
</td></tr> | |||
<tr style="height: 0px;"><td style="border-bottom: solid #000000 1px; border-left: solid #000000 1px; border-right: solid #000000 1px; border-top: solid #000000 1px; padding: 7px 7px 7px 7px; vertical-align: top;"><div dir="ltr" style="line-height: 1.15; margin-bottom: 0pt; margin-top: 0pt; text-align: center;"> | |||
<span style="font-size: x-small;"><span style="background-color: transparent; color: black; font-family: Arial; font-style: normal; font-variant: normal; font-weight: bold; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">Total volume (</span><span style="background-color: white; color: black; font-family: Arial; font-style: normal; font-variant: normal; font-weight: bold; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">µl</span><span style="background-color: transparent; color: black; font-family: Arial; font-style: normal; font-variant: normal; font-weight: bold; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">)</span></span></div> | |||
</td><td style="border-bottom: solid #000000 1px; border-left: solid #000000 1px; border-right: solid #000000 1px; border-top: solid #000000 1px; padding: 7px 7px 7px 7px; vertical-align: top;"><div dir="ltr" style="line-height: 1.15; margin-bottom: 0pt; margin-top: 0pt; text-align: center;"> | |||
<span style="background-color: transparent; color: black; font-family: Arial; font-size: x-small; font-style: normal; font-variant: normal; font-weight: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">50.0</span></div> | |||
</td><td style="border-bottom: solid #000000 1px; border-left: solid #000000 1px; border-right: solid #000000 1px; border-top: solid #000000 1px; padding: 7px 7px 7px 7px; vertical-align: top;"><div dir="ltr" style="line-height: 1.15; margin-bottom: 0pt; margin-top: 0pt; text-align: center;"> | |||
<span style="background-color: transparent; color: black; font-family: Arial; font-size: x-small; font-style: normal; font-variant: normal; font-weight: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">50.0</span></div> | |||
</td><td style="border-bottom: solid #000000 1px; border-left: solid #000000 1px; border-right: solid #000000 1px; border-top: solid #000000 1px; padding: 7px 7px 7px 7px; vertical-align: top;"><div dir="ltr" style="line-height: 1.15; margin-bottom: 0pt; margin-top: 0pt; text-align: center;"> | |||
<span style="background-color: transparent; color: black; font-family: Arial; font-size: x-small; font-style: normal; font-variant: normal; font-weight: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">50.0</span></div> | |||
</td><td style="border-bottom: solid #000000 1px; border-left: solid #000000 1px; border-right: solid #000000 1px; border-top: solid #000000 1px; padding: 7px 7px 7px 7px; vertical-align: top;"><div dir="ltr" style="line-height: 1.15; margin-bottom: 0pt; margin-top: 0pt; text-align: center;"> | |||
<span style="background-color: transparent; color: black; font-family: Arial; font-size: x-small; font-style: normal; font-variant: normal; font-weight: normal; text-decoration: none; vertical-align: baseline; white-space: pre-wrap;">50.0</span></div> | |||
</td></tr> | |||
</table> | |||
</div> | |||
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Revision as of 23:18, 5 June 2014
 iGEM Project name 1
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Thursday, 5th of June 2014
Make 10 mM ATP stock for 1 ml by doing serial dilution.
Make the recipe for ligation. Each person in the team will use 4 micro centrifuge tube to create the control, 1mM ATP, 2mM ATP, and 4 mM ATP. The amount of insert and backbone added varies due to the different amount of concentration of nucleic acid in each set. The ratio of insert to backbone must be 3:1. And since the length of the insert and backbone is 1 kb to 2 kb respectively, the amount of insert must be 30 ng and backbone must be 20 ng. ( total amount of both insert and backbone is ~50 ng)
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