Difference between revisions of "Team:SPSingapore/Notebook-Week-2"

Line 156: Line 156:
  
 
<br><br>
 
<br><br>
 +
 +
<span class = "blue"> Invasin + Listerolysin</span>
 +
&emsp;&emsp;
 +
<font style = "color:blue;font-weight:bold;font-size:11pt;">&#9728; Yan Ting &#9728;</font>
 +
<br><br>
 +
<div class = "divnbcontent">
 +
<span class = "nbcontent">
 +
RE with XbaI and PstI
 +
<br>&emsp;&emsp;&emsp;&emsp; RE - 2ul
 +
<br>&emsp;&emsp;&emsp;&emsp; Buffer - 5ul
 +
<br>&emsp;&emsp;&emsp;&emsp; DNA (4x of miniprep=74.5ng/ul) - 13.4ul
 +
<br>&emsp;&emsp;&emsp;&emsp; H2O - 29.6ul
 +
<br>&emsp;&emsp;&emsp;&emsp; Incubate at 37degC for 2.5h (1130-1400)
 +
</span>
 +
<br><br>
 +
<span>
 +
&#9654; Results: Gel loaded 1kb ladder, uncut, RE digested. 100V for 1h.
 +
<br> Plasmid doesn't have XbaI site - RE digested DNA is a linear 6kb band.
 +
</span>
 +
 +
</div>
 +
 +
<br><br>
 +
  
 
</div>
 
</div>

Revision as of 19:59, 17 September 2015


Research Notebook

Week 16 (6/9 - 12/9)

▪ May 31

Invasin + Listerolysin    esa Quorum Sensing
☀ Xin Yi ☀

Sent EsaR clone 4 and INv-4 for sequencing.


Invasin + Listerolysin    ☀ Yan Ting ☀

RE with XbaI and PstI
     RE - 2ul
     Buffer - 5ul
     DNA (4x of miniprep=74.5ng/ul) - 13.4ul
     H2O - 29.6ul
     Incubate at 37degC for 2.5h (1130-1400)

▶ Results: Gel loaded 1kb ladder, uncut, RE digested. 100V for 1h.
Plasmid doesn't have XbaI site - RE digested DNA is a linear 6kb band.


▪ June 2

Anaerobic Promoter    ☀ Xin Yi ☀

YFP and GFP transformation results - no colonies for YFP
The GFP transformation repeated with 100ng of plasmid was sucessful. 4 colonies of gfp plasmid were inoculated in 3mL LB+amp and grown overnight.


▪ June 3

Anaerobic Promoter    ☀ Yi Han ☀

Miniprep of gfp plasmids
RE digest with EcoRI and RsaI


▪ June 4

Anaerobic Promoter    ☀ Chi Yan ☀

RE digest indicated a very faint smaller band for colonies 2-4, and hence these were likely to be positive clones


▪ June 5

Invasin + Listerolysin    esa Quorum Sensing
☀ Yan Ting ☀

Inoculated 3mLS of Inv-4 and EsaR-4 plasmid carrying bacteria into 100mL LB+ appropriate Antibiotic
Cell culture-> HEK293 cells revived from freezing down appeared detached.


▪ June 6

Invasin + Listerolysin    esa Quorum Sensing    Anaerobic Promoter
☀ Yi Han ☀

Storage of bacterial glycerol stocks for Inv-4, EsaR4 in 25% glycerol
Yanting: grew HEK293T cells in T25 flask
Gel extract to clean up gfp plasmids which loading dye had accidentally been added to.