Difference between revisions of "Will Notebook2"
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==[[User:Wideloache|Wideloache]] 15:45, 15 January 2009 (EST)== | ==[[User:Wideloache|Wideloache]] 15:45, 15 January 2009 (EST)== | ||
I ran my digestion of J61002-J23100 (E/P) on a 0.8% gel and got the following result: | I ran my digestion of J61002-J23100 (E/P) on a 0.8% gel and got the following result: | ||
| − | [[Image:2009-01-15 14hr 27min.jpg|thumb|none| | + | [[Image:2009-01-15 14hr 27min.jpg|thumb|none|75px|0.8% gel. Lane 1: J61002-J23100 (E/P)]] |
I expected to see a band at around 900, which would have been the RFP insert, but since the vector was fairly faint as well, it probably just wasn't bright enough to see. I excised the band at around 2000 bps which should be pSB1A2 vector cut with E/P. | I expected to see a band at around 900, which would have been the RFP insert, but since the vector was fairly faint as well, it probably just wasn't bright enough to see. I excised the band at around 2000 bps which should be pSB1A2 vector cut with E/P. | ||
| − | I also ran my digestions of | + | I also ran my digestions of LacIX86-pLux' and LacII12X86-pLuxLas on a 2.0% gel and got the following result. |
[[Image:2009-01-15 14hr 28min.jpg|thumb|none|140px|2.0% gel. Lane 1: LacIX86-pLux' (E/P). Lane 2: LacII12X86-pLuxLas (E/P)]] | [[Image:2009-01-15 14hr 28min.jpg|thumb|none|140px|2.0% gel. Lane 1: LacIX86-pLux' (E/P). Lane 2: LacII12X86-pLuxLas (E/P)]] | ||
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==[[User:Wideloache|Wideloache]] 20:36, 13 January 2009 (EST)== | ==[[User:Wideloache|Wideloache]] 20:36, 13 January 2009 (EST)== | ||
| − | Today was my first day back in the lab of the semester. The first thing that I needed to do was to get the two pLux promoters that had been synthesized by GeneArt into pSB1A2. I had done an (E/P) digestion of pLux' and pLuxLas last semester that seemed to be successful. However, when I tried to transform into pSB1A2, I had a low colony yield. I decided to make a fresh stock of pSB1A2 digested with E/P. I used the part J61002-J23100. Cutting this with E/P woudl result in a pSB1A2 vector with a RFP expression insert. In this way I could screen out unsuccessful ligations. In addition to running this digestion O/N, I also | + | Today was my first day back in the lab of the semester. The first thing that I needed to do was to get the two pLux promoters that had been synthesized by GeneArt into pSB1A2. I had done an (E/P) digestion of pLux' and pLuxLas last semester that seemed to be successful. However, when I tried to transform into pSB1A2, I had a low colony yield. I decided to make a fresh stock of pSB1A2 digested with E/P. I used the part J61002-J23100. Cutting this with E/P woudl result in a pSB1A2 vector with a RFP expression insert. In this way I could screen out unsuccessful ligations. In addition to running this digestion O/N, I also digested pLux' and pLuxLas with E/P from the minipreps of LacIX86-pLux' and LacII12X86-pLuxLas. |
<pre> | <pre> | ||
Revision as of 20:50, 15 January 2009
Wideloache 15:45, 15 January 2009 (EST)
I ran my digestion of J61002-J23100 (E/P) on a 0.8% gel and got the following result:
I expected to see a band at around 900, which would have been the RFP insert, but since the vector was fairly faint as well, it probably just wasn't bright enough to see. I excised the band at around 2000 bps which should be pSB1A2 vector cut with E/P.
I also ran my digestions of LacIX86-pLux' and LacII12X86-pLuxLas on a 2.0% gel and got the following result.
This was what I expected to see, and I excised the lowest band on each gel (expected size was 93 bps)
Wideloache 20:36, 13 January 2009 (EST)
Today was my first day back in the lab of the semester. The first thing that I needed to do was to get the two pLux promoters that had been synthesized by GeneArt into pSB1A2. I had done an (E/P) digestion of pLux' and pLuxLas last semester that seemed to be successful. However, when I tried to transform into pSB1A2, I had a low colony yield. I decided to make a fresh stock of pSB1A2 digested with E/P. I used the part J61002-J23100. Cutting this with E/P woudl result in a pSB1A2 vector with a RFP expression insert. In this way I could screen out unsuccessful ligations. In addition to running this digestion O/N, I also digested pLux' and pLuxLas with E/P from the minipreps of LacIX86-pLux' and LacII12X86-pLuxLas.
Make GenR plates (15 ug/mL) Miniprep pLas' and pLasLux Get GFP plasmid to put 4 promoters in front of.
