Minnesota/17 July 2008

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(Difference between revisions)
Line 24: Line 24:
!| Parts !! 10x Buffer !! BSA !! H20 !! DNA !! RE 1 !! RE 2
!| Parts !! 10x Buffer !! BSA !! H20 !! DNA !! RE 1 !! RE 2
|-
|-
-
|L2a, LAMBDAcI:Term ||5.0uL ||0.5uL ||32.5uL ||10.0uL ||EcoRI ||Xba1
+
|L2a, LAMBDAcI:Term ||5.0uL ||0.5uL ||32.5uL ||10.0uL ||EcoRI || Xba1
|-
|-
-
|L2b, LAMBDAcI:Term ||5.0uL ||0.5uL ||32.5uL ||10.0uL ||EcoRI ||Xba1
+
|L2b, LAMBDAcI:Term ||5.0uL ||0.5uL ||32.5uL ||10.0uL ||EcoRI || Xba1
|-
|-
-
|L3i, Pro:LAMBDAcI ||5.0uL ||0.5uL ||32.5uL ||10.0uL ||Pst1 ||Spe1
+
|L3i, Pro:LAMBDAcI ||5.0uL ||0.5uL ||32.5uL ||10.0uL || Pst1 || Spe1
|-
|-
-
|L3j, Pro:LAMBDAcI ||5.0uL ||0.5uL ||32.5uL ||10.0uL ||Pst1 ||Spe1
+
|L3j, Pro:LAMBDAcI ||5.0uL ||0.5uL ||32.5uL ||10.0uL || Pst1 || Spe1
|-
|-
-
|L4b, LAMBDAcI:Term ||5.0uL ||0.5uL ||40.5uL ||2.0uL ||EcoRI ||Xba1
+
|L4b, LAMBDAcI:Term ||5.0uL ||0.5uL ||40.5uL || 2.0uL || EcoRI || Xba1
|-
|-
-
|L4c, LAMBDAcI:Term ||5.0uL ||0.5uL ||39.5uL ||3.0uL ||EcoRI ||Xba1
+
|L4c, LAMBDAcI:Term ||5.0uL ||0.5uL ||39.5uL || 3.0uL || EcoRI || Xba1
|-
|-
-
|L4d, LAMBDAcI:Term ||5.0uL ||0.5uL ||39.5uL ||3.0uL ||EcoRI ||Xba1
+
|L4d, LAMBDAcI:Term ||5.0uL ||0.5uL ||39.5uL || 3.0uL || EcoRI || Xba1
|-
|-
-
|L4e, LAMBDAcI:Term ||5.0uL ||0.5uL ||39.5uL ||3.0uL ||EcoRI ||Xba1
+
|L4e, LAMBDAcI:Term ||5.0uL ||0.5uL ||39.5uL || 3.0uL || EcoRI || Xba1
|-
|-
-
|L4g, LAMBDAcI:Term ||5.0uL ||0.5uL ||39.5uL ||3.0uL ||EcoRI ||Xba1
+
|L4g, LAMBDAcI:Term ||5.0uL ||0.5uL ||39.5uL ||3.0uL || EcoRI || Xba1
|-
|-
-
|L4h, LAMBDAcI:Term ||5.0uL ||0.5uL ||39.5uL ||3.0uL ||EcoRI ||Xba1
+
|L4h, LAMBDAcI:Term ||5.0uL ||0.5uL ||39.5uL ||3.0uL || EcoRI || Xba1
|-
|-
-
|L4i, LAMBDAcI:Term ||5.0uL ||0.5uL ||37.5uL ||5.0uL ||EcoRI ||Xba1
+
|L4i, LAMBDAcI:Term ||5.0uL ||0.5uL ||37.5uL ||5.0uL || EcoRI || Xba1
|-
|-
-
|L4j, LAMBDAcI:Term ||5.0uL ||0.5uL ||32.5uL ||10.0uL ||EcoRI ||Xba1
+
|L4j, LAMBDAcI:Term ||5.0uL ||0.5uL ||32.5uL ||10.0uL || EcoRI || Xba1
|-
|-
 +
|L5c, GFP:Term ||5.0uL ||0.5uL ||40.5uL ||2.0uL || EcorI || Xba1
 +
|-
 +
|L6a, Pro:LAMBDAcI:Term ||5.0uL ||0.5uL ||39.5uL ||3.0uL || EcoRI || Xba1
 +
|-
 +
|L6b, Pro:LAMBDAcI:Term ||5.0uL ||0.5uL ||41.5uL || 1.0uL ||EcoRI || Xba1

Revision as of 19:29, 17 July 2008

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1. Plasmid prep dual promoters: (1) Tet & P22mnt, (2) LacI & LAMBDAcI.
2. Model: Figure out why have such a low GFP output when run model.
3. Sequence L5, L6-L9
4. Re-transform RFP, TetR promoter, terminator because no cell growth on plates.
5. Discuss problems with sequences
6. Double digest of all ligations: from previous days. Do a double digest to cut the components linearly, which will then be able to run through a gel to check if correct DNA is present. Follow the table below:
Parts 10x Buffer BSA H20 DNA RE 1 RE 2
L2a, LAMBDAcI:Term 5.0uL 0.5uL 32.5uL 10.0uL EcoRI Xba1
L2b, LAMBDAcI:Term 5.0uL 0.5uL 32.5uL 10.0uL EcoRI Xba1
L3i, Pro:LAMBDAcI 5.0uL 0.5uL 32.5uL 10.0uL Pst1 Spe1
L3j, Pro:LAMBDAcI 5.0uL 0.5uL 32.5uL 10.0uL Pst1 Spe1
L4b, LAMBDAcI:Term 5.0uL 0.5uL 40.5uL 2.0uL EcoRI Xba1
L4c, LAMBDAcI:Term 5.0uL 0.5uL 39.5uL 3.0uL EcoRI Xba1
L4d, LAMBDAcI:Term 5.0uL 0.5uL 39.5uL 3.0uL EcoRI Xba1
L4e, LAMBDAcI:Term 5.0uL 0.5uL 39.5uL 3.0uL EcoRI Xba1
L4g, LAMBDAcI:Term 5.0uL 0.5uL 39.5uL 3.0uL EcoRI Xba1
L4h, LAMBDAcI:Term 5.0uL 0.5uL 39.5uL 3.0uL EcoRI Xba1
L4i, LAMBDAcI:Term 5.0uL 0.5uL 37.5uL 5.0uL EcoRI Xba1
L4j, LAMBDAcI:Term 5.0uL 0.5uL 32.5uL 10.0uL EcoRI Xba1
L5c, GFP:Term 5.0uL 0.5uL 40.5uL 2.0uL EcorI Xba1
L6a, Pro:LAMBDAcI:Term 5.0uL 0.5uL 39.5uL 3.0uL EcoRI Xba1
L6b, Pro:LAMBDAcI:Term 5.0uL 0.5uL 41.5uL 1.0uL EcoRI Xba1
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