907
edits
Changes
added notes about verifying clones
* Incubate for 10 min at RT
* Transform 2uL of this into competent cells (see [[Transformation of Bacteria]]), plating the entire transformation.
===Verification===
* Miniprep the transformed clones and digest 1 ug of purified plasmid for 1h with AgeI and BbsI in NEB Buffer 1.1 and then run on an agarose gel. Empty vectors will yield a ~1kb fragment, wheras vectors with insert will only be cut once. This is because the cloning removes the BbsI site.
* Send clones with insert for sequencing with the hU6 sequencing primer. Align those clones with the empty vector full sequence or the hU6 sequencing of the empty vector, available at http://www.addgene.org/42335/sequences/. Doublecheck that your insert is correct, and is oriented in the correct manner.
