Difference between revisions of "Part:BBa K733001:Design"

Latest revision as of 04:53, 24 September 2012

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)

@@ Line 1: / Line 1: @@
+===Design Note===
-__NOTOC__
+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.
-<partinfo>BBa_K733001 short</partinfo>
-<partinfo>BBa_K733001 SequenceAndFeatures</partinfo>
+===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.
-===Design Notes===
+The sequences of these two primers are:
-Not available at this moment.
+Forward primer: 5’-CATGAAGTCTCCTTGAAATCAGAAGATATTTAGGATATATTTTTCTATGGAT–3’(Prefix not shown)
+Reverse primer: 5’–CAATATCCCTTTTATCCATAGAAAAATATATCCTAAATATCT–3’ (Suffix not shown)
-===Source===
-Not available at this moment.
 ===References===