Versions Compared

Old Version 1

changes.mady.by.user Unknown User (eersland@mit.edu)

Saved on

compared with

New Version Current

changes.mady.by.user Unknown User (eersland@mit.edu)

Saved on

Key

  • This line was added.
  • This line was removed.
  • Formatting was changed.

Results

 
tube nameHyper Ladder 1kbGG DONRpENTR Lyn-mKate 1pENTR Lyn-mKate 1pENTR Lyn-mKate 2pENTR Lyn-mKate 3pENTR Lyn-mKate 4pENTR Lyn-TEV 1pENTR Lyn-TEV 1pENTR Lyn-TEV 2pENTR Lyn-TEV 3pENTR Lyn-TEV 4pENTR Lyn Stop 1pENTR Lyn Stop 1pENTR Lyn Stop 2pENTR Lyn Stop 3pENTR Lyn Stop 4Hyper Ladder 1Hyper Ladder 1GG DONRpENTR Syk-eYFP 1pENTR Syk-eYFP 1pENTR Syk-eYFP 2pENTR Syk-eYFP 3pENTR Syk-eYFP 4ggDONRpENTR CD79B Gal4VP16 1pENTR CD79B Gal4VP16 1pENTR CD79B Gal4VP16 2pENTR CD79B Gal4VP16 3pENTR CD79B Gal4VP16 4pENTR Gmab Heavy 1pENTR Gmab Heavy 1pENTR Gmab Heavy 2pENTR Gmab Heavy 3pENTR Gmab Heavy 4pZDonor 3-GTW-4pEXPR hef1a:CD79B-TCS-Gal4VP16 1pEXPR hef1a:CD79B-TCS-Gal4VP16 1pEXPR hef1a:CD79B-TCS-Gal4VP16 2pEXPR hef1a:CD79B-TCS-Gal4VP16 3Hyper Ladder 1
gel/lane123456789101112131415161718212223242526272829303132333435363738
DNA mass desired (ng)500500500500500500
total volume(ul)202020202020
DNA concentration (ng/ul)  13213214211039
Enzyme Name PvuIIApaIundigestedPvuIIApaIPvuIIApaIPvuIIPvuIIundigestedPvuIIPvuIIPvuIIPvuIIundigestedPvuIIPvuIIPvuIIApaI
Buffer NamePvuII  NsiIundigested controlNsiINsiINsiINsiIPciIundigestedPciIPciIPciIPciIundigestedPciIPciIPciIPciI result             FP?FF    FFFP?  PPPP PPPP 
notes              looks like there are two bands really close together, which there should be         middle band seems slightly shorter than it should be            
tube nameHyper Ladder 1ggDONRpENTR CD79B stop 1pENTR CD79B stop 1pENTR CD79B stop 2pENTR CD79B stop 3pENTR CD79B stop 4pZDoner 3-GTW-4pEXPR 3-hef1a:CD79A-4 1pEXPR 3-hef1a:CD79A-4 1pEXPR 3-hef1a:CD79A-4 2pEXPR 3-hef1a:CD79A-4 3pEXPR 3-hef1a:CD79A-4 4pEXPR 3-hef1a:CD79A -tcs-gal4vp16-4 1pEXPR 3-hef1a:CD79A -tcs-gal4vp16-4 1pEXPR 3-hef1a:CD79A -tcs-gal4vp16-4 2pEXPR 3-hef1a:CD79A -tcs-gal4vp16-4 3pEXPR 3-hef1a:CD79A -tcs-gal4vp16-4 4Hyper Ladder 1Hyper Ladder 1ggDONRpENTR Gmab Light 1pENTR Gmab Light 1pENTR Gmab Light 2pENTR Gmab Light 3pENTR Gmab Light 4pZDonner 2-GTW-3pEXPR 2-TRE:Syk-TEV-3 1pEXPR 2-TRE:Syk-TEV-3 1pEXPR 2-TRE:Syk-TEV-3 2pEXPR 2-TRE:Syk-TEV-3 3pEXPR 2-TRE:Syk-TEV-3 4pEXPR 2-TRE:Syk-3 1pEXPR 2-TRE:Syk-3 1pEXPR 2-TRE:Syk-3 2pEXPR 2-TRE:Syk-3 3pEXPR 2-TRE:Syk-3 4Hyper Ladder 1
gel/lane4142434445464748495051525354555657585961626364656667686970717273747576777879
Enzyme Name PciI/KpnIPciI/KpnIPciI/KpnIPciI/KpnIPciI/KpnIPciI/KpnIKpnIundigestedKpnIKpnIKpnIKpnIundigestedKpnIKpnIKpnIKpnI  NsiIundigestedNsiINsiINsiINsiIBamHIundigestedBamHIBamHIBamHIBamHIundigestedBamHIBamHIBamHIBamHI 
result   FP?P?P?  PPPP PPPP    PFPP  PPPP PPFP 
gel/lane123456
DNA volume#DIV/0!#DIV/0!3.793.793.524.55
buffer volume222222
enzyme volume111234
water volume#DIV/0!#DIV/0!13.212.211.59.5
gel/lane123456
loading dye444567
result      
notes couldn't findnotes    doesn't look like control, but there should be a band around 200                         

 

      

 

Image RemovedImage RemovedImage Removed

Virtual gels from Geneious

fail pZDoner

pEXPR hEF1a: CD79B-TCS-Gal4VP16

pEXPR hEF1a: CD79B+STOP

pEXPR hEF1a: Gmab Heavy

pEXPR hEF1a: Gmab Light

pZDonor
pENTR Syk-eYFP
pENTR Lyn-TEV
pENTR Lyn+STOP
pENTR Lyn-mKate
pENTR CD79B stoppENTR CD79B Gal4VP16pENTR Gmab HeavypENTR Gmab LightpEXPR 3-hef1a:CD79A -4pEXPR 3-hef1a:CD79A-tcs-gal4vp16 -4pEXPR 2-TRE:Syk-TEV-3pEXPR 2-TRE:Syk-3
Fail state (ggDONR)
Fail state (ggDONR QQ ligation) 3-GTW-4fail pZDonner 2-GTW-3
Image RemovedImage RemovedImage RemovedImage RemovedImage RemovedImage RemovedImage RemovedImage RemovedImage RemovedImage RemovedImage RemovedImage Removed

 

 

 

Image Removed

 

Image RemovedImage RemovedImage Removed
 
Image AddedImage AddedImage AddedImage AddedImage Added