User:Karmella Haynes/Notebook/BioBrick cloning/2010/04/20

From OpenWetWare
Jump to: navigation, search
Owwnotebook icon.png Project name Report.pngMain project page
Resultset previous.pngPrevious entry      Next entryResultset next.png

04/20/10

  • ✓ Digests: check plasmids KAH156, 157/MV2; KAH96, 142, 146, 147/V0201



Digests
--> NEB AvaI (buffer 4) for KAH156, 157/MV2 (AvaI cuts 1x at end of insert and 1x in fshPcCD of KAH157)
--> Fermentas FD XbaI for KAH96, 142, 146, 147/V0201


Reagent 1, 2 3-6 Expected:
1. KAH156/MV2 = 5739, 352, 192
2. KAH157/MV2 = 5423, 352, 316, 192
3. KAH96/V0201 = 2825, 5157
4. KAH142/V0201 = 4571, 5157
5. KAH146/V0201 = 6386, 5157
6. KAH147/V0201 = 8132, 5157
File:KAH 042110 gel1.tif
15 μL/lane; 1% agarose
DNA(plasmid) 2.0 2.0
10X buffer 1.5 1.5
AvaI 2.0 ---
XbaI --- 1.0
dH2O 9.5 10.5
  15 μL

--> 37°C/ ~1 hour


--> Unsure about KAH156, 157/MV2; re-submit for sequencing