Minnesota/9 July 2008

From 2008.igem.org

(Difference between revisions)

Revision as of 20:45, 9 July 2008

Back to Notebook Home
Go to Previous Day (July 8)	Go to Next Day (July 10)

1. Single Digests: Performed today. Incubated digested samples for 2hrs @ 37C. Refer to table below.

Gene	10x Buffer	BSA Protein	H20	DNA	RE
Promoter	5.0uL	0.5uL	40.4uL	2.1uL	2.0uL, SPE
LAMBDAcI	5.0uL	0.5uL	29.5uL	15.0uL	1.0uL, XBA 1
LAMBDAcI	5.0uL	0.5uL	29.5uL	15.0uL	1.0uL, SPE
Terminator	5.0uL	0.5uL	38.0uL	4.5uL	2.0uL, XBA 1

NOTE: RE = Restriction Enzyme

2. Ligation: Performed today. Heat inactivated enzyme @ 65C for 15 min to kill enzymes.Ligate samples. Incubate ligated samples @ 16C for 1hour. Refer to table below.

Genes	10x Buffer	H20	Base Vector	Insert	DNA Ligase
LAMBDAcI/Terminator	2.0uL	10.7uL	2.0uL (Term.)	4.3uL (LAMBDAcI)	1.0uL
Promoter/LAMBDAcI	2.0uL	10.7uL	2.0uL (Pro.)	4.3uL (LAMBDAcI)	1.0uL

3. Double Digests

Gene	10x Buffer	BSA Protein	H20	DNA	RE 1	RE 2
Promoter/LAMBDAcI	5.0uL	0.5uL	22.5uL	20.0uL	1.0uL, EcorI	1.0uL, PstI
LAMBDAcI/Terminator	5.0uL	0.5uL	20.0uL	20.0uL	1.0uL, EcorI	1.0uL, PstI
Base Vector	5.0uL	0.5uL	15.0uL	15.0uL	1.0uL, EcorI	1.0uL, PstI

4. Ligation

5. Transformation: Transform ligation products.

6. Make/Pour Ampicillin Plates

7. Order Primers for Part Sequencing

8. Purified Plasmid Prep: Purified MCherry, Promoter, Terminator and Base Vector because sequencing results were poor.

@@ Line 65: / Line 65: @@
 |-
 |7. '''Order Primers for Part Sequencing'''
+|-
+|8. '''Purified Plasmid Prep''': Purified MCherry, Promoter, Terminator and Base Vector because sequencing results were poor.
 |}