Team:The University of Alberta/24 July 2008

From 2008.igem.org

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(General)
 
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*Did mini-preps of J61003 because we were running out
*Did mini-preps of J61003 because we were running out
*Digested BisDA/B with single digests using AgeI and NgoMIV to test the enzymes because weve been getting cruddy digests when using both of them
*Digested BisDA/B with single digests using AgeI and NgoMIV to test the enzymes because weve been getting cruddy digests when using both of them
 +
**'''UPDATE''': It would appear that AgeI is cutting but NgoMIV isn't. Could be a buffer issue?
 +
==System 2==
==System 2==
Today we have taken the ligation from yesterday and transformed it as well as digested it.  
Today we have taken the ligation from yesterday and transformed it as well as digested it.  
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**Then ligated the B0034+TETR with LacIQERE giving us a complete system 2.
**Then ligated the B0034+TETR with LacIQERE giving us a complete system 2.
**PCRed the digested product i.e tetR+BOO34
**PCRed the digested product i.e tetR+BOO34
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*Digested pTet with Spe/Pst.
+
*Digested pTet with Spe/Pst. Column purified the digested product.
*Digested RFP with Xba/Pst.
*Digested RFP with Xba/Pst.
==System 3==
==System 3==
*Digested 5ul of J61003 with Xba/Pst; this will let us insert the Tryp biobrick digested and ligated yesterday
*Digested 5ul of J61003 with Xba/Pst; this will let us insert the Tryp biobrick digested and ligated yesterday
**Ran the digested J6 on a gel; gel purified
**Ran the digested J6 on a gel; gel purified

Latest revision as of 21:20, 24 July 2008

General

  • Did mini-preps of J61003 because we were running out
  • Digested BisDA/B with single digests using AgeI and NgoMIV to test the enzymes because weve been getting cruddy digests when using both of them
    • UPDATE: It would appear that AgeI is cutting but NgoMIV isn't. Could be a buffer issue?

System 2

Today we have taken the ligation from yesterday and transformed it as well as digested it.

  • if all goes well the digested product should be B0034+TETR
    • Column Purified the digested product.
    • Then ligated the B0034+TETR with LacIQERE giving us a complete system 2.
    • PCRed the digested product i.e tetR+BOO34
  • Digested pTet with Spe/Pst. Column purified the digested product.
  • Digested RFP with Xba/Pst.

System 3

  • Digested 5ul of J61003 with Xba/Pst; this will let us insert the Tryp biobrick digested and ligated yesterday
    • Ran the digested J6 on a gel; gel purified