Part:BBa_K1041002:Experience

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Team NRP-UEA_Norwich 2013

Team NRP-UEA_Norwich 2013 created this part using biobricks BBa_K1041000 and BBa_K1041001. These biobricks both contain a Nde1 site after their promoter sequence, enabling a restriction digest to be performed. The RFP coding gene was excised from BBa_K1041000 and ligated in front of the AntG promoter of BBa_K1041001 to create a new biobrick.

Characterisation of this biobrick involved sequencing, restriction digests and BLAST analysis.

Restriction Digest

Part Bba_K1041002 was digested with enzymes XbaI and NdeI and compared to uncut DNA Fig 1.

Fig 1:Lane 1 contains and Bba_K1041002 cut with XbaI and NdeI and lane 2 uncut Bba_K1041002.

Sequencing

The biobrick was sent off to a company for sequencing and the data we received back Fig 2,3,4 showed the DNA is good quality.

Fig 2:K1041002 sequencing data part 1

Fig 3:K1041002 sequencing data part 2

Fig 4:K1041002 sequencing data part 3

BLAST Analysis

The data we recieved back from the sequencing company was aligned using BLAST with the expected DNA sequence Fig 5,6

User Reviews

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