Haynes Lab:Notebook/Investigating Photo-Switchable Synthetic Nucleosomes/2012/11/12

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Great! Next step...

Digests:
- In a 30 μL reaction, cut H2B with XbaI and SpeI
- In a 30 μL reaction, cut LOV with XbaI and SpeI
- In a 30 μL reaction, cut a pSB1A2 plasmid (from Rene) XbaI and SpeI

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-==Entry title==
+==November 12, 2012==
-* Insert content here...
+* The PCR products were purified with the Zymo DNA Clean & Concentrator Kit
+{| class="wikitable" border=1 cellpadding="5" cellspacing="0"
+|-
+| PCR Product || 260|| 280 || 260/280 || ng/μL
+|-
+| H2B || 0.052 || 0.028 || 1.865 || 51.607
+|-
+| LOV || 0.048 || 0.026 || 1.860 || 48.095
+|}
+----
+Great! Next step...
+* Digests:
+** In a 30 μL reaction, cut H2B with XbaI and SpeI
+** In a 30 μL reaction, cut LOV with XbaI and SpeI
+** In a 30 μL reaction, cut a [http://partsregistry.org/Part:pSB1A2 pSB1A2 plasmid] (from Rene) XbaI and SpeI
+{| class="wikitable" border=1 cellpadding="5" cellspacing="0"
+|-
+| Reagent || H2B|| LOV || pSB1A2
+|-
+| DNA || 8.0 || 8.0 || 15.0
+|-
+| XbaI || 1.0|| 1.0 || 1.0
+|-
+| SpeI || 1.0 || 1.0 || 1.0
+|-
+| 10x buffer || 3.0 || 3.0 || 3.0
+|-
+| dH<sub>2</sub>O || --- || --- || ---
+|-
+| Total vol. || 30. 0 || 30.0  || 30.0
+|}
+* Run entire 30 μL on a 1% gel (use the big fat-tooth well comb)
+* Cut band out of gel (use UV box); H2B = ~410, LOV = ~461, pSB1A2 = ~2000
+* Extract DNA from gel