1291 pET30b-TAT-NLS-sso7d

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Cloning of 1291 pET30b-TAT-NLS-sso7d

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Purpose: Cloning of 1291 pET30b-TAT-NLS-sso7d

Date: July 31, 2012

Notebook: 

Page:

Clone ID: 1291

Experimenters: Jennifer Truong

Results

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Cloning

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Date: July 31, 2012


Gene synthesis of PCR product. Expected size ~100 bp

See lane 2

Cut out and purify band, digest with RE  Nde1 + Kpn1  (lane 1) with vector 368 pET30b-sso7d-KX (lane 2)

Cut out bands, purify and ligate over night

Date: August 6, 2012

MiniPrep screening came back all negative, started all over again


Gene synthesis of PCR product. Expected size ~100 bp

See lane 2

Cut out and purify band, digest with RE Nde1 + Kpn1 (lane 1) with vector 368 pET30b-sso7d-KX (lane 2)


Cut out bands, purify and ligate over night

Colony Screening

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Digested MiniPrep with RE Nde1 + Kpn1, no inserts

Date: August 9, 2012

Digested MIniPrep with RE Nde1 and Kpn1, faint inserts at expected ~100 bp

1, 2, 3, 5 and 6 may be correct

Result intepretation/Summary

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Conclusion

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