Haynes Lab:Notebook/Engineering PC-TFs/2015/05/07

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Summary

  • Miniprep'd liquid cultures.

All samples between 100-250 ng/μL.


Digest and Gel Purification

Reagent Volume
DNA (2μg) 23μL 12μL 12.0 μL 12.0 μL 20.0 μL 16.0 μL 20.0 μL (From well 2 to well 8)
10X buffer 3.0 μL 3.0 μL 3.0 μL 3.0 μL 3.0 μL 3.0 μL 3.0 μL
XbaI 0μL 0μL 0μL 0μL 2μL 2μL 2μL
PstI 2μL 2μL 2μL 2μL 2.0μL 2μL 2μL
SpeI 2μL 2μL 2μL 2μL 0.0 μL 0μL 0μL
dH2O 0μL 11μL 11μL 11μL 3μL 7μL 7μL
Total 30 μL --> 37°C/ 15 min.

Ladder

Linker kah and pctf gel purification.JPG

15 μL/lane; 1% agarose;
Expected:
All linkers (KAH021-024) [SpeI/PstI]
V0120 + linker (very short). Only one band expected.

Inserts (xPcD-RFP-VP64; KAH0879,KAH089,KAH090 [XbaI/PstI]
1053bp


Observed lengths:
W1: Ladder
W2: KAH021
W3: KAH022
W4: KAH023
W5: KAH024
W6: KAH087
W7: KAH089
W8: KAH090