Team:Yale/Our Project/Notebook/Week 4

From 2010.igem.org

(Difference between revisions)
Line 112: Line 112:
<h4>PCR amplification of phs </h4>  
<h4>PCR amplification of phs </h4>  
<li> Because the gel run the previous day was no longer good, had to run a second 0.8% agarose gel of the PCR products to determine the success of the PCR reaction. Below is the image of the gel, with a 1 kb ladder in the leftmost lane, phsC in next lane, phsAB in lane 3, and phsABC in lane 4. </li>
<li> Because the gel run the previous day was no longer good, had to run a second 0.8% agarose gel of the PCR products to determine the success of the PCR reaction. Below is the image of the gel, with a 1 kb ladder in the leftmost lane, phsC in next lane, phsAB in lane 3, and phsABC in lane 4. </li>
 +
<br />
<div align="center">
<div align="center">
<img src="http://2010.igem.org/wiki/images/6/68/Yale-1st-phs-pcr.jpg" />
<img src="http://2010.igem.org/wiki/images/6/68/Yale-1st-phs-pcr.jpg" />
</div>
</div>
 +
<br />
<div id="caption">
<div id="caption">
-
The phsC fragment seems to have been amplified with decent success, but phsABC and phAB are only present in trace amounts, so the thermocycler protocol will have to be tweaked. <br/> </div>
+
The phsC fragment seems to have been amplified with decent success, but phsABC and phAB <br /> are only present in trace amounts, so the thermocycler protocol will have to be tweaked. <br/> </div>
<li> In the mean time, gel extract the phsC from lane 2 and the phsABC from lane 4 using the microcentrifuge variant of the  <a href="http://2010.igem.org/Team:Yale/Our_Project/Protocols/gel_extract">standard gel extraction protocol </a> </li>
<li> In the mean time, gel extract the phsC from lane 2 and the phsABC from lane 4 using the microcentrifuge variant of the  <a href="http://2010.igem.org/Team:Yale/Our_Project/Protocols/gel_extract">standard gel extraction protocol </a> </li>

Revision as of 22:03, 27 October 2010

iGEM Yale

lab notebook: week 4 (6/28 -7/4)

  • Monday 6/28--PCR amplification of thiosulfate reductase operon phsABC
  • See more/less
  • Tuesday 6/29--Gel extraction of PCR amplified phs, TSI agar slant assay for hydrogen sulfide production, & pre-ligation double digestion
  • See more/less
  • Wednesday 6/30--Troubleshooting PCR amplification of thiosulfate reductase gene & continued work toward ligation
  • See more/less
  • Thursday 7/1--Smelly bacteria (yay!) & ongoing ligation efforts
  • See more/less
  • Friday 7/2--Confirmation of hydrogen sulfide production plus more ligation work
  • See more/less
  • Saturday 7/3--More attempts to put a terminator on thiosulfate reductase and a promoter on lacI
  • See more/less
  • Sunday 7/4--Ligation attempt #2 transformants
  • See more/less