Team:KIT-Kyoto/Notebook/ATF1/may
From 2013.igem.org
(Created page with "{{Template:KIT-Kyoto-1}} {{Template:KIT-Kyoto-header}} <html class="no-js"> <head> </head> </html> <div class="document"> <div id="doc-left"> {{Template:KIT-Kyoto/menu_note}}...") |
|||
Line 13: | Line 13: | ||
<div id="doc-right" class="metro"> | <div id="doc-right" class="metro"> | ||
+ | <h2>ATF1</h2> | ||
<P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT COLOR="#444444"><FONT SIZE=4><B>May | <P STYLE="margin-bottom: 0cm"><FONT FACE=""><SPAN LANG="en-US"><FONT COLOR="#444444"><FONT SIZE=4><B>May |
Revision as of 10:47, 26 September 2013
ATF1
May 16th
We performed PCR to amplify the ATF1 gene.
Primers:
Forward:5’-TACGCCTCGAGATGAATGAAATCGATGAGAAAAATCAGGCCCC-3’
Reverse:5’-ATATTGCTTAGCAGGGCCTAAAAGGAGAGCTTTGTAAATGGAGCAAAGC-3’
Reaction composition is as follows:
Buffer |
50uL |
dNTP |
20uL |
Primer mix |
1uL |
DNA sample |
0.5uL |
KOD-FX |
2uL |
H2O |
26.5uL |
total |
100uL |
We could not get any PCR product.
May 17th -20th
Changed PCR conditions with another DNA template. We performed PCR of the ATF1 gene again.
Primers:
Forward
Reverse:5’-ATATTGCTTAGCAGGGCCTAAAAGGAGAGCTTTGTAAATGGAGCAAAGC-3’
Reaction composition is as follows:
Buffer |
50uL |
dNTP |
20uL |
Primer mix |
1uL |
DNA sample |
0.5uL |
KOD-FX |
2uL |
H2O |
26.5uL |
total |
100uL |
We could not get any PCR product.
May 30th -31th
We transformed a plasmid carrying the ATF1 gene into E.coli.
This plasmid was obtained from iGEM DNA distributions kit 2012 (plate 2, o-7).
We could see approximately 50 colonies and cultured furthermore.