Team:Heidelberg/Notebook/BSDesign/October

From 2010.igem.org

(Difference between revisions)
(→10/05/2010)
Line 22: Line 22:
:3= 10S
:3= 10S
:4= 10L
:4= 10L
 +
 +
 +
== 10/06/2010 ==
20 Miniprep:
20 Miniprep:
Line 69: Line 72:
|20||1179||4.8
|20||1179||4.8
|}
|}
 +
 +
* digestion of miMeasure and 6 different synthetic miRNA patterns against endogenous miR122
 +
* gel seperation
 +
* gel purification
 +
* sequencing
----
----
 +
 +
== 10/12/2010 ==
 +
 +
4 miraPCRs for binding sites of miR4286.
 +
* one PCR with an annealing temperature of 45 degree and 16 cycles
 +
* second miraPCR with annealing temperature of 57 degree and also 16 cycles
 +
**nucleotide removal
 +
**digestion
 +
**gel seperation
 +
**gel purification
 +
**ligation in miMeasure and pSB1C3
 +
**transformation
 +
 +
== 10/13/2010 ==
 +
 +
*pick colonies
 +
*about 8 hours later: do minipreps
 +
*send for sequencing
{{:Team:Heidelberg/Single_Bottom}}
{{:Team:Heidelberg/Single_Bottom}}

Revision as of 23:55, 27 October 2010
 


Binding Site Design - October

10/02/2010


  • digestion in 50 µl of
    • 4x pSB1C3 (5µl BSA, 5µl NEB buffer 2, 1µl PstI, 1µl EcoRI, 1µl DpnI, 4µl construct, 33 µl H2O)
    • 4x pSB1A3 (5µl BSA, 5µl NEB buffer 2, 1µl PstI, 1µl EcoRI, 1µl DpnI, 4µl construct, 33 µl H2O)

10/05/2010


  • 1µg digestion of miMeasure and 6 different constructs (1A, 3F, 13, 15, 17, 37) with EcoRI and PstI
1= 0S
2= 0L
3= 10S
4= 10L


10/06/2010

20 Miniprep:

Table 1: 20 Minis
Numberbinding site against miRNA xPosition
12213.5
22211.4
311793.1
442862.6
542864.1
642863.4
711794.4
83.5
911791.9
102214.6
112211.5
1242863.5
132214.8
142212.3
1542862.2
1642861.3
1711792.5
1811792.4
1911791.5
2011794.8
  • digestion of miMeasure and 6 different synthetic miRNA patterns against endogenous miR122
  • gel seperation
  • gel purification
  • sequencing


10/12/2010

4 miraPCRs for binding sites of miR4286.

  • one PCR with an annealing temperature of 45 degree and 16 cycles
  • second miraPCR with annealing temperature of 57 degree and also 16 cycles
    • nucleotide removal
    • digestion
    • gel seperation
    • gel purification
    • ligation in miMeasure and pSB1C3
    • transformation

10/13/2010

  • pick colonies
  • about 8 hours later: do minipreps
  • send for sequencing

Retrieved from "http://2010.igem.org/Team:Heidelberg/Notebook/BSDesign/October"