IGEM:Peking/2007/Count-Conjugation-Notebook/2007-8-23: Difference between revisions

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(New page: =Lock & Key By Yu Tao= ==Acknowledgement and Farewell== *First, I have to say thank you to Mingzhi because he helped me with almost all the experiments these ten days so that I could spend...)
 
 
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*First, I have to say thank you to Mingzhi because he helped me with almost all the experiments these ten days so that I could spend most of my time accompanying my girlfriend. n_n Thank you Mingzhi.  
*First, I have to say thank you to Mingzhi because he helped me with almost all the experiments these ten days so that I could spend most of my time accompanying my girlfriend. n_n Thank you Mingzhi.  
*Second, I am going home today and I am very excited. Goodbye and Work hard, my friends. I like the feeling of home, Ah Ha! :) I will be back on Sep, 1st.
*Second, I am going home today and I am very excited. Goodbye and Work hard, my friends. I like the feeling of home, Ah Ha! :) I will be back on Sep, 1st.
=Tandem OriT by Mingzhi Qu & Ze Ren=
==Conjugation Test:R751_pSC101 X Dh5α_psb1A2(III)==
*Donor:      C600-R751_psc101 (Tc+)
*Recipient:  Dh5α+ psb1A2 (Amp+)
*Control:
#donnor：C600_R751
#donnor：DH5α_pSC101
*mixing condition: 220rpm, 90min.
*'''Day1:'''
#Get the plates from -4 fridge：C600-R751_psc101(Tc+), C600_psb1A2(Amp+), C600_R751, Dh5α_psc101
#Amplification Culture in liquid LB for 12 hours.
*'''Day2:'''
*put 2mL of culture into 20mL of LB with antibiotics, sub-culturing.
====Donor====
*final OD600:
<pre>
#R751+psc101    0.510
#C600_R751      0.470
#C600_psc101    0.480
</pre>
====Recipient====
*final OD600:
<pre>
#Dh5α+pSB1A2    1.631
</pre>
====Plating the conjugant mix ====
*C600_R751_psc101 group was set up serial dilutions:original, 10-1, 10-2.
*culture on Tc+/Amp+ plate.
====result(Next day)====
<pre>
No Donor                  X  recipient  time rpm  clones(original)  clones(e-1)  clones(e-2)
1  C600_R751_psc101        X  Dh5α_pSB1A2  90  220      full              1000+      <10
2  C600_R751              X  Dh5α_pSB1A2  90  220      40+                /          /
3  C600_psc101            X  Dh5α_pSB1A2  90  220      full              /          / 
</pre>

Latest revision as of 04:33, 2 September 2007

Lock & Key By Yu Tao

Acknowledgement and Farewell

  • First, I have to say thank you to Mingzhi because he helped me with almost all the experiments these ten days so that I could spend most of my time accompanying my girlfriend. n_n Thank you Mingzhi.
  • Second, I am going home today and I am very excited. Goodbye and Work hard, my friends. I like the feeling of home, Ah Ha! :) I will be back on Sep, 1st.

Tandem OriT by Mingzhi Qu & Ze Ren

Conjugation Test:R751_pSC101 X Dh5α_psb1A2(III)

  • Donor: C600-R751_psc101 (Tc+)
  • Recipient: Dh5α+ psb1A2 (Amp+)
  • Control:
  1. donnor：C600_R751
  2. donnor：DH5α_pSC101
  • mixing condition: 220rpm, 90min.
  • Day1:
  1. Get the plates from -4 fridge：C600-R751_psc101(Tc+), C600_psb1A2(Amp+), C600_R751, Dh5α_psc101
  2. Amplification Culture in liquid LB for 12 hours.
  • Day2:
  • put 2mL of culture into 20mL of LB with antibiotics, sub-culturing.

Donor

  • final OD600:
#R751+psc101    0.510
#C600_R751      0.470
#C600_psc101    0.480

Recipient

  • final OD600:
#Dh5α+pSB1A2    1.631

Plating the conjugant mix

  • C600_R751_psc101 group was set up serial dilutions:original, 10-1, 10-2.
  • culture on Tc+/Amp+ plate.

result(Next day)

No Donor                   X   recipient  time rpm  clones(original)  clones(e-1)   clones(e-2)
1  C600_R751_psc101        X  Dh5α_pSB1A2  90  220      full              1000+       <10
2  C600_R751               X  Dh5α_pSB1A2  90  220       40+                /          /
3  C600_psc101             X  Dh5α_pSB1A2  90  220       full               /          /
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