Haynes Lab:Notebook/Engineering PC-TFs/2015/04/13

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Summary

  • Restriction Digest/Gel Verification, Concentration Reading

Digest and Gel Verification

Reagent Volume

Expected:
BL01 Ligation Colonies [XbaI/ApaLI]
Forward insertion: 255, 288, 883, 1243, 2037, 3017
Reverse insertion: 288, 255, 1122, 1243, 2037, 2783
No insertion: 268, 883, 1243, 2783

BL09 Ligation Colonies [XbaI/ApaLI]
Forward insertion: 255, 288, 883, 1243, 3977
Reverse insertion: 255, 288, 1243, 2082, 2783
No insertion: 268, 883, 1243, 2783

4-13-15.jpg

15 μL/lane; 1% agarose;
Observed lengths:
W1: Ladder
W2: Bl09 TL V1
W3: Bl09 TL V2
W4: BL01 LSR V1
W5: BL01 LSR V2
W6: BL01 LSR V3
W7: BL01 LSR V4

Ladder

DNA 3.0 μL 3.0 μL 2.0 μL 2.0 μL 1.0 μL 1.0 μL (From ladder 2 to ladder 6)
10X buffer 1.5.0 μL (for all)
XbaI 1.0 μL (for all)
ApaLI 1.0 μL (for all)
dH2O 8.5 μL 8.5 μL 9.5 μL 9.5 μL 10.5 μL 10.5 μL
Total 15 μL --> 37°C/ 15 min.


Note: Punctured hole in Well 3. Wells 4 & 5 do not match with Wells 6 & 7. Rather, Wells 6 & 7 match with Wells 2 & 3. The tubes may have been mislabeled and Wells 6 & 7 could be Bl09 rather than Bl01. We will do another restriction digest and gel verification later this week to verify if that is the case.