Team:LMU-Munich/Notebook/Pathway

From 2010.igem.org

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mastermix:
mastermix:
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{|
+
::{|
-
|-
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| MQ: 93.6µl
| MQ: 93.6µl
|-
|-
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|-
|-
| Phusion: 1.2µl (Pfu-Promega)
| Phusion: 1.2µl (Pfu-Promega)
 +
|}
 +
::->109.2µl/6 = 18.2µl each tube
 +
 +
----
 +
{| class="wikitable" border="1"
 +
|-
 +
!
 +
! 1
 +
! 2
 +
! 3
 +
! 4
 +
! 5
 +
! 6
 +
|-
 +
| '''primer fwd'''
 +
| pduAfwd (1)
 +
| pduJfwd (7)
 +
| 1P1D (12)
 +
| mpduDfwd (9)
 +
| 5P3AD (14)
 +
| 5P3AD (14)
 +
|-
 +
| '''primer rev'''
 +
| pduJrev (8)
 +
| pduUrev (2)
 +
| mpduDrev (10)
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| 3P2D (13)
 +
| 9P4A (15)
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| 9P4A (15)
 +
|-
 +
| '''template'''
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| Knut
 +
| Knut
 +
| gDNA ''citrobacter freundii''
 +
| gDNA ''c. freundii''
 +
| gDNA ''streptomyces thioluteus'', glycerolstock
 +
| gDNA ''s. thioluteus'', LB prep
|}
|}

Revision as of 09:39, 25 August 2010


Pathway Notebook

Contents

Week Days
Monday Tuesday Wednesday Thursday Friday Saturday Sunday
31 8-02-2010 8-03-2010 8-04-2010 8-05-2010 8-06-2010 8-07-2010 8-08-2010
32 8-09-2010 8-10-2010 8-11-2010 8-12-2010 8-13-2010 8-14-2010 8-15-2010
33 8-16-2010 8-17-2010 8-18-2010 8-19-2010 8-20-2010 8-21-2010 8-22-2010
34 8-23-2010 8-24-2010 8-25-2010 8-26-2010 8-27-2010 8-28-2010 8-29-2010
35 8-30-2010 8-31-2010 9-01-2010 9-02-2010 9-03-2010 9-04-2010 9-05-2010
36 9-06-2010 9-07-2010 9-08-2010 9-09-2010 9-10-2010 9-11-2010 9-12-2010
37 9-13-2010 9-14-2010 9-15-2010 9-16-2010 9-17-2010 9-18-2010 9-19-2010
38 9-20-2010 9-21-2010 9-22-2010 9-23-2010 9-24-2010 9-25-2010 9-26-2010
39 9-27-2010 9-28-2010 9-29-2010 9-30-2010 10-01-2010 10-02-2010 10-03-2010

8-02-2010

  • Some test text here.

8-03-2010

Some test text in bold We created following tests:

  • test1
  • test2
  • test3

8-04-2010

Example of a table

header 1 header 2 header 3
row 1, cell 1 row 1, cell 2 row 1, cell 3
row 2, cell 1 row 2, cell 2 row 2, cell 3


8-05-2010

gDNA prep (cultures from 7-28-2010) with innuPREP Bacteria DNA kit

-> additionally TE-buffer (100µl EDTA [o.5M]; 500µl Tris [1M]) was made, pH=8.0

8-06-2010

text

8-07-2010

weekend

8-08-2010

weekend

8-09-2010

PCR were performed as follows:

mastermix:

MQ: 93.6µl
10xbuffer: 12.0µl
dNTP's: 2.4µl (each 10mM)
Phusion: 1.2µl (Pfu-Promega)
->109.2µl/6 = 18.2µl each tube
1 2 3 4 5 6
primer fwd pduAfwd (1) pduJfwd (7) 1P1D (12) mpduDfwd (9) 5P3AD (14) 5P3AD (14)
primer rev pduJrev (8) pduUrev (2) mpduDrev (10) 3P2D (13) 9P4A (15) 9P4A (15)
template Knut Knut gDNA citrobacter freundii gDNA c. freundii gDNA streptomyces thioluteus, glycerolstock gDNA s. thioluteus, LB prep

8-10-2010

text

8-11-2010

text

8-12-2010

text

8-13-2010

text

8-14-2010

weekend

8-15-2010

weekend

8-16-2010

text

8-17-2010

text

8-18-2010

text

8-19-2010

text

8-20-2010

text

8-21-2010

weekend

8-22-2010

weekend

8-23-2010

text

8-24-2010

Preparing Competent Cells with Benny's protocoll & buffers

- grow the culture to an OD 600 of ~ 0,3

-> our cells: OD 600 = 0,256

- centrifuge for 10 min at 4°C at 3000rpm

- resuspend the pellet in buffer 1

-> we used 400 µl

- centrifuge for 10 min at 4°C at 2500rpm

- resuspend the pellet in less buffer 1 than before

-> we used 200 µl

- add buffer 2 in a ratio of 1:10 = buffer 2:buffer 1

-> we added 20µl of buffer 2

- incubate on ice for 10 min

-> as we probably used too little buffer, we added another 200 µl of buffer 1 and 20µl of buffer 2

- aliquote the suspension and shockfreeze it at -80°C

-> we put 50 µl in each eppendorf


Test-Transformation in order to see whether the sells will work

- done with protocoll (3 Transformation)

-> we tested with the following DNA:

1. K098200 (biobrick) [Amp., 5 µl]
2. pUC [Amp., 1µl]

-> as the cells are probably in a to high concentration, we thined them down with saline to an end-concentration of 10 -2

8-25-2010

text

8-26-2010

text

8-27-2010

text

8-28-2010

weekend

8-29-2010

weekend

8-30-2010

text

8-31-2010

text

9-01-2010

text

9-02-2010

text

9-05-2010

text

9-04-2010

weekend

9-05-2010

weekend

9-06-2010

text

9-07-2010

text

9-08-2010

text

9-09-2010

text

9-10-2010

text

9-11-2010

weekend

9-12-2010

weekend

9-13-2010

text

9-14-2010

text

9-15-2010

text

9-16-2010

text

9-17-2010

text

9-18-2010

weekend

9-19-2010

weekend

9-20-2010

text

9-21-2010

text

9-22-2010

text

9-23-2010

text

9-24-2010

text

9-25-2010

weekend

9-26-2010

weekend

9-27-2010

text

9-28-2010

text

9-29-2010

text

9-30-2010

text

10-01-2010

text

10-02-2010

weekend

10-03-2010

weekend

 

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