hEF1a:CD79AhEF1a:CD79BhEF1a:LynhEF1a:GMabHhEF1a:GMabLhEF1a:CD79A-TCS-G4hEF1a:CD79B-TCS-G4TRE:Syk-15-TEVphEF1a:rtTAhEF1a:Gal4VP16hEF1a-LacO:mKateTRE:mKatehEF1a:LacITRE:Gal4VP16pENTR hEF1a-LacOpENTR LacITRE:Syk
Number1234567891011121314151617
Enzyme

ApaI

Cuts in hEF1a, CD79A & backbone

ApaI

Cuts in hEF1a, and backbone

ApaI, NcoI

Cuts in Lyn, hEF1a and backbone

BglII

Cuts in GmabH, hEF1a and backbone

KpnI

Cuts in GmabL, hEF1a, and backbone

PvuII

Cuts in CD79A, TCS and backbone

PvuII

Cuts in CD79B, TCS and backbone

PvuII

Cuts in syk, tevp and backbone

ApaI

Cuts in hEF1a, rtTA3 and backbone

XhoI

Cuts in hEF1a, Gal4VP16 and backbone

NheIPvuIIApaIXhoI

ApaI

cuts in hEF1a-LacO

ApaI

cuts in LacI

PvuII

cuts in syk

Virtual gel
Concentration (ng/uL)1110.314201314.4983.51359.01086.1487.21161.9706.4441.9741.0483.1420.61265.3229.8196.7321.8
Buffer                 
Enzyme                 
DNA volume0.450328740.3521126760.38040170.5083884090.3679175860.4603627661.0262725780.4303296320.707814271.131477710.6747638331.034982411.188777940.3951632022.1758050482.541942041.55376
H2O volume11.5496712611.6478873211.61959811.4916115911.6320824111.5396372310.9737274211.5696703711.292185710.8685222911.3252361710.965017610.811222111.60483689.8241949529.4580579610.44624
Buffer volume1.51.51.51.51.51.51.51.51.51.51.51.51.51.51.51.51.5
Enzyme volume1.51.51.51.51.51.51.51.51.51.51.51.51.51.51.51.51.5
1-1011-17A-JK-Q

hEF1a:CD79A Correct

hEF1a: CD79B Correct

hEF1a:Lyn Correct

hEF1a: GmabH Correct

hEF1a:GmabL Correct

hEF1a:CD79A-TCS-G4 Correct

hEF1a:CD79B-TCS-G4 I can't tell

TRE:Syk-15-TEVp Correct

hEF1a:rtTA Correct

hEF1a:Gal4VP16 ???

hEF1a-LacO:mKate Correct

TRE:mKate Doesn't match either condition

hEF1a:LacI Correct

TRE:Gal4VP16 Correct

pENTR hEF1a-LacO Correct

pENTR LacI right band pattern but shifted down a bit -> could be a janky gel. Probably correct because hEF1a:LacI

TRE:Syk Correct

All failed golden gatesAll failed golden gates
hEF1a:CD79A-linker-Gal4VP163T33T93T156T66T96T129T39T69T99T129T1512T612T912T1215T315T915T15
LetterABCDEFGHIJKLMNOPQ
Concentration (ng/uL)1838.335.325.131.026.8209.9402.9175.6249.1311.629.227.232.717.619.323.5
DNA volume1212121212122.3820871.2410032.847382.0072261.604621121212121212
H2O volume0000009.61791310.7599.152629.99277410.39538000000
Buffer volume1.51.51.51.51.51.51.51.51.51.51.51.51.51.51.51.51.5
Enzyme volume1.51.51.51.51.51.51.51.51.51.51.51.51.51.51.51.51.5
  • No labels