1121 pET28b-Exo-1

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Cloning of 1121 pET28b-Exo-1

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Purpose: Cloning of 1121 pET28b-Exo-1

Date: July 23, 2012

Notebook: 

Page:

Clone ID: 1121

Experimenters: Jennifer Truong

Results

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Cloning

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Date: July 24, 2012


See lane 2, expected size 1428 bp

PCR from MG1665 genomic DNA

Exo-I-Nde1-F
Nde1
cagacaCAT ATGATGAATGACGGTAAGCAAC

Exo-I-Xho1-R
(GGCAGTACGCGGAAGAGATTGTC CTCGAG cagatc)
gatctgCTCGAGGACAATCTCTTCCGCGTACTGCC

Cut out band, purify, digest with RE Ndel and Xho1, with vector pET28b

See lanes 6 (insert) and 7 (vector), cut out bands, purify and ligate together and transform

Colony Screening

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Date: July 27, 2012

Digested MiniPrep with RE Nde1 and Xho1, see lanes 13-18

All samples are positive for insert, submitted sample 2 for sequencing

Result intepretation/Summary

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Conclusion

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