TUDelft/22 August 2008

From 2008.igem.org

July
MTWTFSS
  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
MTWTFSS
        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
MTWTFSS
[http://2008.igem.org/TUDelft/1_September_2008 1] [http://2008.igem.org/TUDelft/2_September_2008 2] [http://2008.igem.org/TUDelft/3_September_2008 3] [http://2008.igem.org/TUDelft/4_September_2008 4] [http://2008.igem.org/TUDelft/5_September_2008 5] [http://2008.igem.org/wiki/index.php?title=TUDelft/6_September_2008&action=edit 6] [http://2008.igem.org/wiki/index.php?title=TUDelft/7_September_2008&action=edit 7]
[http://2008.igem.org/TUDelft/8_September_2008 8] [http://2008.igem.org/TUDelft/9_September_2008 9] [http://2008.igem.org/TUDelft/10_September_2008 10] [http://2008.igem.org/TUDelft/11_September_2008 11] [http://2008.igem.org/TUDelft/12_September_2008 12] [http://2008.igem.org/wiki/index.php?title=TUDelft/13_September_2008&action=edit 13] [http://2008.igem.org/wiki/index.php?title=TUDelft/14_September_2008&action=edit 14]
[http://2008.igem.org/TUDelft/15_September_2008 15] [http://2008.igem.org/TUDelft/16_September_2008 16] [http://2008.igem.org/TUDelft/17_September_2008 17] [http://2008.igem.org/TUDelft/18_September_2008 18] [http://2008.igem.org/TUDelft/19_September_2008 19] [http://2008.igem.org/wiki/index.php?title=TUDelft/20_September_2008&action=edit 20] [http://2008.igem.org/wiki/index.php?title=TUDelft/21_September_2008&action=edit 21]
[http://2008.igem.org/TUDelft/22_September_2008 22] [http://2008.igem.org/TUDelft/23_September_2008 23] [http://2008.igem.org/TUDelft/24_September_2008 24] [http://2008.igem.org/TUDelft/25_September_2008 25] [http://2008.igem.org/wiki/index.php?title=TUDelft/26_September_2008&action=edit 26] [http://2008.igem.org/wiki/index.php?title=TUDelft/27_September_2008&action=edit 27] [http://2008.igem.org/wiki/index.php?title=TUDelft/28_September_2008&action=edit 28]
[http://2008.igem.org/TUDelft/29_September_2008 29] [http://2008.igem.org/TUDelft/30_September_2008 30]
October
MTWTFSS
    [http://2008.igem.org/TUDelft/1_October_2008 1] [http://2008.igem.org/TUDelft/2_October_2008 2] [http://2008.igem.org/TUDelft/3_October_2008 3] [http://2008.igem.org/wiki/index.php?title=TUDelft/4_October_2008&action=edit 4] [http://2008.igem.org/wiki/index.php?title=TUDelft/5_October_2008&action=edit 5]
[http://2008.igem.org/TUDelft/6_October_2008 6] [http://2008.igem.org/TUDelft/7_October_2008 7] [http://2008.igem.org/TUDelft/8_October_2008 8] [http://2008.igem.org/TUDelft/9_October_2008 9] [http://2008.igem.org/TUDelft/10_October_2008 10] [http://2008.igem.org/wiki/index.php?title=TUDelft/11_October_2008&action=edit 11] [http://2008.igem.org/wiki/index.php?title=TUDelft/12_October_2008&action=edit 12]
[http://2008.igem.org/TUDelft/13_October_2008 13] [http://2008.igem.org/TUDelft/14_October_2008 14] [http://2008.igem.org/TUDelft/15_October_2008 15] [http://2008.igem.org/TUDelft/16_October_2008 16] [http://2008.igem.org/TUDelft/17_October_2008 17] [http://2008.igem.org/wiki/index.php?title=TUDelft/18_October_2008&action=edit 18] [http://2008.igem.org/wiki/index.php?title=TUDelft/19_October_2008&action=edit 19]
[http://2008.igem.org/TUDelft/20_October_2008 20] [http://2008.igem.org/TUDelft/21_October_2008 21] [http://2008.igem.org/TUDelft/22_October_2008 22] [http://2008.igem.org/TUDelft/23_October_2008 23] [http://2008.igem.org/TUDelft/24_October_2008 24] [http://2008.igem.org/wiki/index.php?title=TUDelft/25_October_2008&action=edit 25] [http://2008.igem.org/wiki/index.php?title=TUDelft/26_October_2008&action=edit 26]
[http://2008.igem.org/wiki/index.php?title=TUDelft/27_October_2008&action=edit 27] [http://2008.igem.org/wiki/index.php?title=TUDelft/28_October_2008&action=edit 28] [http://2008.igem.org/wiki/index.php?title=TUDelft/29_October_2008&action=edit 29] [http://2008.igem.org/wiki/index.php?title=TUDelft/30_October_2008&action=edit 30] [http://2008.igem.org/wiki/index.php?title=TUDelft/31_October_2008&action=edit 31]

August 22nd

DNA extraction test

Today another gel was run with DNA soaked from the spots. To test the soaking-protocol, various steps of the protocol were performed on the DNA. This time 10 ul of TE was used, because due to the soaking of the water into the punch, using 5 ul reduced free liquid to be added to cells (or for analysis) below 2 ul. Again various conditions were tested, and also the later sent spots from the promoter strength testing DNA were analyzed. To give an idea of DNA concentrations, all spots were nanodropped. These results are in the table below. For this testing we used a spot we won't be using and showed a good band on the gel of the repository. 1002 9A, a lock from Berkeley06, which had inconsistent sequence.

Nanodrop of extracted DNA
Sample ng/ul A260/A280
TE (blank) 0.53 1.42
10 ul TE + 1002 9A punch, Tr 4.94 0.69
10 ul TE + 1002 9A punch, Tr + spinned 9.69 0.89
10 ul TE + 1002 9A punch, T = 42ºC 11.04 0.75
10 ul TE + 1002 9A punch, T = 42ºC + Spinned 11.78 1.23
10 ul TE + promoter strength punch, Tr 5.94 0.97
10 ul TE + promoter strength punch, 42 C + spinned 4.81 1.36
10 ul TE + 3 ul pSB1A7 (isolated on 20-08-08) 11.69 1.66

Tr = Room Temperature


These results have again rather low 260/280, but DNA concentrations that should be visible on a gel. Concentrations this time with increased following of iGEM procedure.

The gel showed no visible DNA bands for the soaked punches. The positive control did show bands, as can be seen below.

TUDelft 220808 extraction.jpg

Transformation

A transformation has been started today for testing of the midiprepped vectors. To prepare for the arrival of our thermosensitive sequences next week we also tried to transform a couple of parts for our constructs, although if these fail we will need a different way to obtain the parts. The parts (apart from the midiprepped) used for a transformation are:

Parts transformed and nanodrop of concentration
Part ng/ul A260/A280
I712019 14.22 0.64
B0015 14.48 0.68
I13453 10.16 0.88
R0010 4.44 0.66
B0030 11.33 0.67
I742152 20.2 0.69


Punches were no longer obtained by the punch tool but by cutting them out with a scalpel, due to time gain and the punch tool not working very well. All these punches were soaked in 10 ul TE and transformations were started with 5 ul of extracted DNA. After transformation cells were plated on LB+Amp.