Team:UNIPV-Pavia/Notebook/Week4Jul

From 2009.igem.org

Revision as of 02:44, 23 July 2009 by Lor18 (Talk | contribs)

EthanolPVanimation.gif

December 2008
M T W T F S S
1 2 3 4 5 6 7
8 9 10 11 12 13 14
15 16 17 18 19 20 21
22 23 24 25 26 27 28
29 30 31
March 2009
M T W T F S S
1
2 3 4 5 6 7 8
9 10 11 12 13 14 15
16 17 18 19 20 21 22
23 24 25 26 27 28 29
30 31
April 2009
M T W T F S S
1 2 3 4 5
6 7 8 9 10 11 12
13 14 15 16 17 18 19
20 21 22 23 24 25 26
27 28 29 30
May 2009
M T W T F S S
1 2 3
4 5 6 7 8 9 10
11 12 13 14 15 16 17
18 19 20 21 22 23 24
25 26 27 28 29 30 31
June 2009
M T W T F S S
1 2 3 4 5 6 7
8 9 10 11 12 13 14
15 16 17 18 19 20 21
22 23 24 25 26 27 28
29 30
July 2009
M T W T F S S
1 2 3 4 5
6 7 8 9 10 11 12
13 14 15 16 17 18 19
20 21 22 23 24 25 26
27 28 29 30 31
August 2009
M T W T F S S
1 2
3 4 5 6 7 8 9
10 11 12 13 14 15 16
17 18 19 20 21 22 23
24 25 26 27 28 29 30
31
September 2009
M T W T F S S
1 2 3 4 5 6
7 8 9 10 11 12 13
14 15 16 17 18 19 20
21 22 23 24 25 26 27
28 29 30
October 2009
M T W T F S S
1 2 3 4
5 6 7 8 9 10 11
12 13 14 15 16 17 18
19 20 21 22 23 24 25
26 27 28 29 30 31
November 2009
M T W T F S S
1
2 3 4 5 6 7 8
9 10 11 12 13 14 15
16 17 18 19 20 21 22
23 24 25 26 27 28 29
30

Week from July 20th, to July 26th, 2009

Previous Week Next Week

July, 20th

  • Overnight digestion (20 ul reaction volume) for:
MRGENE1(X-P) MRGENE1-2(X-P) MRGENE1-3(X-P)
MRGENE2(X-P)(X2) MRGENE2-2(X-P) MRGENE2-3(X-P)
B0030(S-P)(X3)
  • NOTE: reactions which involve excision(X-P) were performed on 3 ug of DNA, while S-P digestions were performed with 1 ug of DNA. We decided to have 3 replicates to be sure to have an acceptable yield.
  • We decided to perform gel run/cut/purification for MRGENE2 and for B0030, while we decided to perform DNA precipitation with sodium acetate for MRGENE1.



  • We transformed about 2.5 pg of A14-4 in TOP10 in order to filter the wanted plasmids. We plated the transformed bacteria and incubated the plates at 37°C overnight.



  • We infected 5 ml of LB + Amp with 10 ul of A12-2 and A12-3 glycerol stocks. Tomorrow they will be miniprepped and sent to sequence!



Preparation of experiment with Tecan F200

  • We infected 5 ml of LB + Kan with a single colony taken from the native plate of B0015.
  • We incubated the inoculum for 6 hours (37°C, 220 rpm).



Experiment with Tecan F200

  • Description
  • Purpose:
  • Materials & Methods
  • Protocol
  • Results


Top

July, 21st

  • Gel run for:
    • MRGENE1(for check)
    • MRGENE2
    • MRGENE2-2
    • MRGENE2-3
    • B0030(X3)
  • We noticed unwanted bands in MRGENE1 run...So, we performed an analysis on Mr Gene plasmids and we found an unwanted PstI site in pMK-RQ that gives the noticed bands...
  • We decided to proceed to ligation and to perform a massive screening of the ligated inserts in the next days.
  • Gel purification for:
    • MRGENE2
    • MRGENE2-2
    • MRGENE2-3
    • B0030(X3)
  • DNA precipitation with sodium acetate for MRGENE1, MRGENE1-2, MRGENE1-3.
  • Results: after quantifications at Nanodrop, all the purified DNA samples had a good yield! let's proceed to ligation!
  • Ligation:
    • B1 = B0030(S-P) + MRGENE1(X-P)
    • B2 = B0030(S-P) + MRGENE2(X-P)
  • We incubated the ligations at 16°C overnight.

Top

July, 22nd

Top


July, 23rd

Top


July, 24th

Top



Previous Week Next Week