Team:BYU Provo/Team Thermosensor/Week6

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==Week 6 (May 22 - 28)==
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* May 23 LacZ ran on gel-product YES!
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** LacZ PCR cleanup
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*** Cleaned up LacZ according to Qiaquick protocol and labeled as T 20-1(needs to be checked on gel)
 +
* May 25
 +
** Restriction Digest of LacZ and GFP
 +
*** Digested GFP at the HindIII and XbaI sites
 +
**** oops...we realized that our reverse LacZ primer had a BglII site in it when we needed an XbaI site. Dr. Grose ordered a new primer with an XbaI restriction site in it.
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** Confirming pBAD clones by restriction digest
 +
*** 5uL buffer 4
 +
*** 0.5 uL BSA
 +
*** 30 uL plasmid(pIG 17-4 and 17-5 each in individual tubes)
 +
*** 12uL H2O
 +
*** THEN MIX WELL AND ADD
 +
*** 1.5 uL BamHI-HF
 +
*** 1.5 uL EcoRI-HF
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**** If the gel shows two bands then pBAD was inserted successfully into pIG17-4 and/or pIG17-5. The Digest of pIG174 will be labeled as RD 20-4 and the digest of pIG17-5 will be labeled RD 20-5.
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**** placed RDs in 37oC bath overnight and placed in the freezer
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* May 27
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**Running RD 20-4 and RD 20-5 on an agarose gel. We will be using a digested pBAD gene and pPLAT digested at BamHI and PstI as our controls on the gel.
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Revision as of 02:31, 24 September 2011

Team BYU Provo

BYU Provo
 

Week 6 (May 22 - 28)

  • May 23 LacZ ran on gel-product YES!
    • LacZ PCR cleanup
      • Cleaned up LacZ according to Qiaquick protocol and labeled as T 20-1(needs to be checked on gel)
  • May 25
    • Restriction Digest of LacZ and GFP
      • Digested GFP at the HindIII and XbaI sites
        • oops...we realized that our reverse LacZ primer had a BglII site in it when we needed an XbaI site. Dr. Grose ordered a new primer with an XbaI restriction site in it.
    • Confirming pBAD clones by restriction digest
      • 5uL buffer 4
      • 0.5 uL BSA
      • 30 uL plasmid(pIG 17-4 and 17-5 each in individual tubes)
      • 12uL H2O
      • THEN MIX WELL AND ADD
      • 1.5 uL BamHI-HF
      • 1.5 uL EcoRI-HF
        • If the gel shows two bands then pBAD was inserted successfully into pIG17-4 and/or pIG17-5. The Digest of pIG174 will be labeled as RD 20-4 and the digest of pIG17-5 will be labeled RD 20-5.
        • placed RDs in 37oC bath overnight and placed in the freezer
  • May 27
    • Running RD 20-4 and RD 20-5 on an agarose gel. We will be using a digested pBAD gene and pPLAT digested at BamHI and PstI as our controls on the gel.
Retrieved from "http://2011.igem.org/Team:BYU_Provo/Team_Thermosensor/Week6"