Team:Freiburg/Notebook/6 September

From 2011.igem.org

(Difference between revisions)
(→=Sequence analysis)
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sequence analysis of our lovtap-Notgate was neagtive:-( So we´ll try the gibson assembly.
sequence analysis of our lovtap-Notgate was neagtive:-( So we´ll try the gibson assembly.
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===PCR===
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'''Investigators: Sophie'''
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'''PCR'''
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{| style="border-spacing:0;"
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| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| Name: Sophie
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| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| Date: 06.09.11
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|-
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| colspan="2"  style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| Continue from Experiment (Date)
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(Name)
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|-
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| colspan="2"  style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| Project Name: Gibson of ♥ and NOT
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|}
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PCR-Mixture for one Reaction:
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For a 50 µl reaction use
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{| style="border-spacing:0;"
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| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| 32,5µl
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| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| H<sub>2</sub>0
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| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| Name
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|-
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| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| 10µl
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| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| 5x Phusion Buffer
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| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| of Primer
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|-
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| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| 2.5µl
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| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| Primer fw
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| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| ♥: P97
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NOT: P99
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|-
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| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| 2.5µl
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| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| Primer dw
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| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| ♥: P98
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NOT: P100
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|-
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| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| 1µl
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| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| dNTPs
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| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| of Template DNA
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|-
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| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| 1µl
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| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| DNA-Template
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| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| NoT, M45 (BBa_Q0400)
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original DNA from Edinburgh (BBa_K322999)
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|-
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| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| 0.5 µl
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| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| Phusion (add in the end)
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| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"|
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|}
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What program do you use?
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First 20 cycles touchdown 69°C -0.4/cycle
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next 10 cycles touchdown 72°C -0.2 per cycle
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To confirm the PCR-Product has the correct size, load 2 µl of the sample onto an agarose-gel.
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How did you label the PCR-Product, where is it stored and what do you do next?
==<span style="color:red;">red light receptor</span>==
==<span style="color:red;">red light receptor</span>==

Revision as of 14:12, 6 September 2011


This is the wiki page
of the Freiburger student
team competing for iGEM 2011.
Thank you for your interest!

Contents

Commons

Miniprep

Investigators: Sandra

Yesterday LB was inoculated with strains (S14, S39-S44 etc.). Miniprep of strains.


Digestion: 2A-assembly

Investigators: Sandra

Digestion of:

  • S14 (GFP)

with XbaI and PstI

  • S39 (PR1)
  • S40 (PR2)
  • S41 (PR3)
  • S42 (PR4)
  • S43 (PR5)
  • S44 (PR6)

with SpeI and PstI

incubation for 6 hours at 37°C.

green light receptor

NAME OF YOUR EXPERIMENT

Investigators:NAME


blue light receptor

Trafo

Investigators: Sandra

Trafo of ligated parts of the 2A-assembly.

Sequence analysis

sequence analysis of our lovtap-Notgate was neagtive:-( So we´ll try the gibson assembly.

PCR

Investigators: Sophie PCR


Name: Sophie


Date: 06.09.11
Continue from Experiment (Date)

(Name)

Project Name: Gibson of ♥ and NOT

PCR-Mixture for one Reaction:

For a 50 µl reaction use


32,5µl H20 Name
10µl 5x Phusion Buffer of Primer
2.5µl Primer fw ♥: P97

NOT: P99

2.5µl Primer dw ♥: P98

NOT: P100

1µl dNTPs of Template DNA
1µl DNA-Template NoT, M45 (BBa_Q0400)

original DNA from Edinburgh (BBa_K322999)

0.5 µl Phusion (add in the end)

What program do you use?

First 20 cycles touchdown 69°C -0.4/cycle

next 10 cycles touchdown 72°C -0.2 per cycle


To confirm the PCR-Product has the correct size, load 2 µl of the sample onto an agarose-gel.


How did you label the PCR-Product, where is it stored and what do you do next?

red light receptor

NAME OF YOUR EXPERIMENT

Investigators:NAME



Lysis cassette

NAME OF YOUR EXPERIMENT

Investigators:NAME


Precipitator

NAME OF YOUR EXPERIMENT

Investigators: NAME

                       

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