Team:Freiburg/Notebook/27 July

From 2011.igem.org

(Difference between revisions)
(→PCR of Lov-tap and Not-gate)
(→PCR of Lov-tap and Not-gate)
Line 15: Line 15:
We repeated the PCR again, with higher temperatures this time.
We repeated the PCR again, with higher temperatures this time.
After PCR we digested with DpnI 37°C for 1 hour and then purified the DNA with PCR purifictaion kit. Afterwards we measured the DNA concentration and loaded 2 microl to a gel.
After PCR we digested with DpnI 37°C for 1 hour and then purified the DNA with PCR purifictaion kit. Afterwards we measured the DNA concentration and loaded 2 microl to a gel.
 +
'''PCR'''
 +
 +
 +
{| style="border-spacing:0;"
 +
| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| Name:Sandra, Sophie
 +
 +
 +
 +
| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| Date: 27.07.11
 +
 +
|-
 +
| 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: NOT-Gate and LovTAP with Gibson-overhangs (now we use a little bit higher temperature for primer annealing) (Date): 25.07.11
 +
 +
(Name): Blue Light
 +
 +
|-
 +
| colspan="2"  style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| Project Name:
 +
 +
|}
 +
PCR-Mixture for one Reaction:
 +
 +
For a 50 µl reaction use
 +
 +
 +
{| style="border-spacing:0;"
 +
| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| 32,5µl
 +
| 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
 +
| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| Name
 +
 +
|-
 +
| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| 10µl
 +
| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| 5x Phusion Buffer
 +
| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| of Primer
 +
 +
|-
 +
| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| 2.5µl
 +
| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| Primer fw
 +
| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| NOT_G_up
 +
 +
LOV_G_up
 +
 +
|-
 +
| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| 2.5µl
 +
| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| Primer dw
 +
| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| NOT_G_dw
 +
 +
LOV_G_dw
 +
 +
|-
 +
| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| 1µl
 +
| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| dNTPs
 +
| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| of Template DNA
 +
 +
|-
 +
| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| 1µl
 +
| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| DNA-Template
 +
| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| S35 (BBa_K322999)
 +
 +
S45 (Bba_Q04400)
 +
 +
|-
 +
| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"| 0.5 µl
 +
| 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)
 +
| style="border:0.0069in solid #00000a;padding-top:0in;padding-bottom:0in;padding-left:0.075in;padding-right:0.075in;"|
 +
 +
|}
 +
What program do you use?
 +
 +
57c auf 70c (in reality: 58c and then 68c)
 +
 +
 +
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?
==<span style="color:red;">red light receptor</span>==
==<span style="color:red;">red light receptor</span>==

Revision as of 13:06, 27 July 2011

Contents

green light receptor

NAME OF YOUR EXPERIMENT

Investigators:NAME


blue light receptor

PCR of Lov-tap and Not-gate

Investigators: Sandra, Sophie

We repeated the PCR again, with higher temperatures this time. After PCR we digested with DpnI 37°C for 1 hour and then purified the DNA with PCR purifictaion kit. Afterwards we measured the DNA concentration and loaded 2 microl to a gel. PCR


Name:Sandra, Sophie


Date: 27.07.11
Continue from Experiment: NOT-Gate and LovTAP with Gibson-overhangs (now we use a little bit higher temperature for primer annealing) (Date): 25.07.11

(Name): Blue Light

Project Name:

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 NOT_G_up

LOV_G_up

2.5µl Primer dw NOT_G_dw

LOV_G_dw

1µl dNTPs of Template DNA
1µl DNA-Template S35 (BBa_K322999)

S45 (Bba_Q04400)

0.5 µl Phusion (add in the end)

What program do you use?

57c auf 70c (in reality: 58c and then 68c)


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

Retrieved from "http://2011.igem.org/Team:Freiburg/Notebook/27_July"