XB-FEAT-6041599 and In situ checklist (Conlon lab): Difference between pages
imported>Xenbase gene generator No edit summary |
imported>Virgilio (Created page with "Protocol checklist submitted by VGP from Conlon lab protocols [http://www.unc.edu/~fconlon/Protocols.htm#tropprot] link to protocol checklist word document [http://www.unc.edu/...") |
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Protocol checklist submitted by VGP from Conlon lab protocols [http://www.unc.edu/~fconlon/Protocols.htm#tropprot] | |||
link to protocol checklist word document [http://www.unc.edu/~fconlon/Protocols/In_Situ_Checklist.doc] | |||
Date:__________ Species:_________ Probes:______________________________ | |||
Day 1 | |||
0.5 vol DEPC 5min RT ___ ___ ___ | |||
PBST 5min RT ___ ___ ___ ___ ___ | |||
Proteinase K 15min RT ___ | |||
*De-vitellinization 10min ___ | |||
PBST 5min RT ___ ___ ___ | |||
Triethanolamine 5min RT ___ ___(4ml) | |||
Acetic Acid Anhydride 5min RT ___ ___ | |||
PBST 5min RT ___ ___ | |||
4% PFA 20min RT ___ | |||
PBST 5min RT ___ ___ ___ ___ ___ | |||
20%Hyb/80%PBST 5min RT ___ | |||
Hyb Buffer o/n 60 ___ | |||
Day 2 | |||
Probe(s): | |||
Day 3 | |||
Hyb Buffer 10min 60 ___ | |||
2xSSC 20min 60 ___ ___ ___ ___ | |||
0.2xSSC 60min 60 ___ repeat if necessary ___ | |||
1xMAB 15min 60 ___ | |||
1xMAB 15min RT ___ | |||
1xMAB/2%BMBR 60min RT ___ | |||
Blocking Buffer 60min RT ___ | |||
Antibody o/n 4 ___ | |||
Day 4 | |||
1xMAB 60min RT ___ ___ ___ ___ | |||
1xMAB o/n RT ___ | |||
Day 5 | |||
AP Buffer 5min RT ___ ___ | |||
BM Purple AP Substrate 1-8hrs RT/4 ___ | |||
PBS 5min RT ___ ___ ___ | |||
Fix:_________________ 30min RT ___ ___ | |||
MeOH 60min RT ___ (stored ___) | |||
Day 6 | |||
Bleaching _____ RT/4 | |||
Latest revision as of 08:52, 18 November 2011
Protocol checklist submitted by VGP from Conlon lab protocols [1]
link to protocol checklist word document [2]
Date:__________ Species:_________ Probes:______________________________
Day 1
0.5 vol DEPC 5min RT ___ ___ ___
PBST 5min RT ___ ___ ___ ___ ___
Proteinase K 15min RT ___
*De-vitellinization 10min ___
PBST 5min RT ___ ___ ___
Triethanolamine 5min RT ___ ___(4ml)
Acetic Acid Anhydride 5min RT ___ ___
PBST 5min RT ___ ___
4% PFA 20min RT ___
PBST 5min RT ___ ___ ___ ___ ___
20%Hyb/80%PBST 5min RT ___
Hyb Buffer o/n 60 ___
Day 2
Probe(s):
Day 3
Hyb Buffer 10min 60 ___
2xSSC 20min 60 ___ ___ ___ ___
0.2xSSC 60min 60 ___ repeat if necessary ___
1xMAB 15min 60 ___
1xMAB 15min RT ___
1xMAB/2%BMBR 60min RT ___
Blocking Buffer 60min RT ___
Antibody o/n 4 ___
Day 4
1xMAB 60min RT ___ ___ ___ ___
1xMAB o/n RT ___
Day 5
AP Buffer 5min RT ___ ___
BM Purple AP Substrate 1-8hrs RT/4 ___
PBS 5min RT ___ ___ ___
Fix:_________________ 30min RT ___ ___
MeOH 60min RT ___ (stored ___)
Day 6
Bleaching _____ RT/4