1290 pET28b-PTD-DRBD
Cloning of 1290 pET28b-PTD-DRBD
[edit]Purpose: Cloning of 1290 pET28b-PTD-DRBD
Date: August 1, 2012
Notebook:
Page:
Clone ID: 1290
Experimenters: Jennifer Truong
Results
[edit]Cloning
[edit]Date: August 1, 2012
<br> [[Image<br> :1292 1307DIGEST10 31 2012.JPG]]<br>
Synthesized template via gene synthesis
Expected size ~400 bp (see lane 1)
Cut out band and purified
Digested the insert (lane 1) and the vector (V12, pET28b) (lane 2) with RE N<br> de1 + Xho1
Cut out bands, pu
rify and ligate together overnight
Date: August 2, 2012
Ligation went wrong yesterday, start all over
Synthesized template via gene synthesis
Expected size ~400 bp (see lane 1)
Cut out band and purified
<br>
Digested the insert (lane 1) and the vector (V12, pET28b) (lane 2) with RE Nde1 + Xho1
Cut out bands, purify and ligate together overnight
Date: August 13, 2012
Sequencing came back with a lot of mutations
Synthesized template via gene synthesis
Expected size ~400 bp (see lane 1)
Cut out band and purified
<br>
Digested the insert (lane 1) and the vector (V12, pET28b) (lane 2) with RE Nde1 + Xho1
Cut out bands, purify and ligate together overnight
Colony Screening
[edit]Date: August 7, 2012
Digested MiniPrep with RE Nde1 and Xho1
Expected size ~400 bp
Samples 1, 2, 3, 5 and 6 may be correct
Sent sample 5 to sequencing
Date: August 16, 2012
Digested MiniPrep with RE Nde1 and Xho1
Expected size ~400 bp
Samples 1, 2 and 6 may be correct. Send sample 6 to sequencing
Date: OCO 26,
2012 Digested insert and vector with Nde1 + Xho1
Ligate , transform
Colony Screening
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Result intepretation/Summary
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Conclusion
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