Latest revision as of 01:48, 24 August 2008

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Things we did today

Grace

Plasmid prep	DNA concentration	260/280	260/230
nir+rbs	394.63 ng/μl	2.04	2.36
rbs+GFP	94.9 ng/μl	2.06	2.17

Grace

File:082308REdigests.jpg

EtBr stained 2% agarose gel ran at 60V for 3 hours. Thirty microliters of RE digest reaction were loaded into each well.

History is the only laboratory we have in which to test the consequences of thought. - Étienne Gilson

[http://manoa.hawaii.edu/ ][http://manoa.hawaii.edu/ovcrge/ ][http://www.ctahr.hawaii.edu ]

@@ Line 7: / Line 7: @@
 :* Plasmid prep of nir+rbs, rbs+GFP
-:* Overnight RE digest of plasmid preps, nir plasmid, GFP plasmid with XbaI and SpeI
+{|class=wikitable border=1 align=center
+!Plasmid prep
+!DNA concentration
+!260/280
+!260/230
+|-
+|nir+rbs
+| 394.63 ng/&mu;l
+|2.04
+|2.36
+|-
+|rbs+GFP
+|94.9 ng/&mu;l
+|2.06
+|2.17
+|}
+:* PCR of insert of nir+rbs, nir (control), rbs+GFP, GFP (control)
 ===3A assembly===
 :<strong> Grace</strong>
-[[Image:082308REdigests.jpg|right|thumb|250px|EtBr stained 2% agarose gel ran at 60V for 3 hours.]]
+[[Image:082308REdigests.jpg|right|thumb|250px|EtBr stained 2% agarose gel ran at 60V for 3 hours. Thirty microliters of RE digest reaction were loaded into each well.]]
 :* Gel purified RE digests
+::* nir band did not show up on gel
 :* Ligated with pSB1A2:
-::* nir+rbs
 ::* plac+rbs
-::* rbs+GFP
 ::* GFP+tt
 ::* GFPf+tt
 :* Transformed DB3.1 cells with 5&mu;l ligation reaction
-===BB-pRL1383a MCS re-replacement===
-:<strong>Grace</strong>
-:* Gel purified RE digests
-:* Ligated J33207 with BB-pRL1383a
-:* Transformed DH5&alpha cells with 5 &mu;l ligation reaction
 = Discussion =