Minnesota/1 August 2008

From 2008.igem.org

(Difference between revisions)
 
Line 17: Line 17:
|-
|-
|'''3. Digest Rxn:''' Digested Terminators, Lac/LAMBDAcI, and GFP. Follow the table below:
|'''3. Digest Rxn:''' Digested Terminators, Lac/LAMBDAcI, and GFP. Follow the table below:
 +
 +
 +
{|border="1" align="left"
 +
|-
 +
!| Parts !! 10x Buffer !! BSA !! H20 !! DNA !! RE 1 !! RE 2 !! Total
 +
|-
 +
| Term 2 || 5.0uL || 0.5uL || 39.5uL || 3.0uL || 1.0uL, Xba1 || 1.0uL, Pst1 || 50.0uL
 +
|-
 +
| Term 4 || 5.0uL || 0.5uL || 28.5uL || 14.0uL || 1.0uL, Xba1 || 1.0uL, Pst1 || 50.0uL
 +
|-
 +
| Lac/LAMBDA A || 5.0uL || 0.5uL || 8.0uL || 34.5uL || 1.0uL, Spe1 || 1.0uL, Pst1 || 50.0uL
 +
|-
 +
| Lac/LAMBDA B || 5.0uL || 0.5uL || 16.0uL || 26.5uL || 1.0uL, Spe1 || 1.0uL, Pst1 || 50.0uL
 +
|-
 +
| GFP || 5.0uL || 0.5uL || 40.5uL || 2.0uL || 1.0uL, Xba1 || 1.0uL, Pst1 || 50.0uL
 +
|}

Latest revision as of 20:11, 4 August 2008

Back to Notebook Home
Go to Previous Day (July 31)Go to Next Day (August 4)
1. Ran the terminator digest out on a gel: After which we were unable to see a band the appropriate length, so we simply cut out the gel where we expected the band to be (terminator is very small - so hard to see). Gel purified - then used this in our next ligation. When spec'ed the purified gel terminator product, there were 2.5ngram/uL of DNA concentration.
2. Currently have two parts that are base vector-promoter-gene, and one that is base vector-promoter: We are currently ligating the terminator to the two parts that have vector-promoter-gene.
The base vector-promoter-gene constructs are: TetR promoter:Lambda cI and Tet/p22 promoter:RFP.
The base vector-promoter construct is: Lambda cI/Lac promoter
3. Digest Rxn: Digested Terminators, Lac/LAMBDAcI, and GFP. Follow the table below:


Parts 10x Buffer BSA H20 DNA RE 1 RE 2 Total
Term 2 5.0uL 0.5uL 39.5uL 3.0uL 1.0uL, Xba1 1.0uL, Pst1 50.0uL
Term 4 5.0uL 0.5uL 28.5uL 14.0uL 1.0uL, Xba1 1.0uL, Pst1 50.0uL
Lac/LAMBDA A 5.0uL 0.5uL 8.0uL 34.5uL 1.0uL, Spe1 1.0uL, Pst1 50.0uL
Lac/LAMBDA B 5.0uL 0.5uL 16.0uL 26.5uL 1.0uL, Spe1 1.0uL, Pst1 50.0uL
GFP 5.0uL 0.5uL 40.5uL 2.0uL 1.0uL, Xba1 1.0uL, Pst1 50.0uL
Retrieved from "http://2008.igem.org/Minnesota/1_August_2008"