Difference between revisions of "Part:BBa K784023"

Revision as of 18:49, 24 September 2012

MCS in pSB1C3

The sequence below has been created in order to expand the number of restriction sites available for shipping BioBricks in pSB1C3. If you want to ship a sequence that can't be digested with restriction sites found in the BioBrick prefix or suffix, this MCS can be the solution.

The addition of the MCS to the pSB1C3 backbone resulted in a change in the BioBrick prefix (one point mutation downstream to the XbaI site). This was in order to add the BglII restriction site in overlap to the XbaI restriction site from the prefix.

The restriction sites available in the MCS are: BglII, HindIII, PacI and BamHI. These restriction sites are preceded by the EcoRI, NotI and XbaI sites from the BioBrick prefix. They are also followed by the SpeI, NotI and PstI sites from the BioBrick suffix.

Usage and Biology

When doing a double digest in the MCS, take into account the required distance between two restriction sites. Keep in mind that each enzyme will cut separately. Once the first enzyme cuts, the rules for [http://www.neb.com/nebecomm/tech_reference/restriction_enzymes/cleavage_olignucleotides.asp#.UFbVTK7kqO4 cleavage close to the end of DNA fragments] from NEB apply. This means, that the distance between two restriction sites should be at least the required distance for each of restriction enzymes when it is close to the end of a DNA fragment.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
INCOMPATIBLE WITH RFC[21]
Illegal BamHI site found at 22
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
COMPATIBLE WITH RFC[1000]

@@ Line 4: / Line 4: @@
 The sequence below has been created in order to expand the number of restriction sites available for shipping BioBricks in [[Part:pSB1C3|pSB1C3]]. If you want to ship a sequence that can't be digested with restriction sites found in the BioBrick prefix or suffix, this MCS can be the solution.
-The addition of the MCS to the [[Part:pSB1C3|pSB1C3]] backbone resulted in a change in the BioBrick prefix. This was in order to add the BglII restriction site in overlap to the XbaI restriction site from the prefix.
+The addition of the MCS to the [[Part:pSB1C3|pSB1C3]] backbone resulted in a change in the BioBrick prefix (one point mutation downstream to the XbaI site). This was in order to add the BglII restriction site in overlap to the XbaI restriction site from the prefix.
 The restriction sites available in the MCS are: BglII, HindIII, PacI and BamHI. These restriction sites are preceded by the EcoRI, NotI and XbaI sites from the BioBrick prefix. They are also followed by the SpeI, NotI and PstI sites from the BioBrick suffix.