Regulatory

Part:BBa_K733001:Design

Designed by: GU, Bida; SUN Fei   Group: iGEM12_HKUST-Hong_Kong   (2012-09-16)
Revision as of 04:53, 24 September 2012 by Feisun0718 (Talk | contribs) (→‎Source)

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Design Note

In order to ease the difficulties we may encounter in amplifying the promoter out directly from B. subtilis genomic DNA, we design two primers mentioned below. Even when doing the PCR with the two primers, it was hard for us to get pure DNA at the first place. Thus, we tried to run for the thermal cycle for only one time, and use phenol chloroform to purify the DNA.

Source

We first obtain the sequence of this part from http://dbtbs.hgc.jp/. Then we design two single strand oligonucleotides (primers) and use only one round of denature, annealing and extension to get our intended promoter.

The sequences of these two primers are:

Forward primer: 5’-CATGAAGTCTCCTTGAAATCAGAAGATATTTAGGATATATTTTTCTATGGAT–3’(Prefix not shown)

Reverse primer: 5’–CAATATCCCTTTTATCCATAGAAAAATATATCCTAAATATCT–3’ (Suffix not shown)

References