RADIOLABELING OF PROBES FOR ELECTROPHORETIC MOBILITY SHIFT ASSAYS
Materials:
g-32 P ATP (New England Nμclear/Perkin Elmer CAT# NEG-035C)
“Μpper strand” oligonμcleotide (at 0 ... [阅读全文]
John Mundy Institute of Molecular BiologyCopenhagenDenmark
http://www.arabidopsis.org/info/Protocols_Mundy2.jsp#Footprint
1)probe: same as used for gelshift)isolated by isotacelectrophoresis w/out E ... [阅读全文]
Sugden labMcArdle Laboratory for Cancer ResearchUniversity of Wisconsin-Madison Medical School DNA /EMSA.pdf">http://mcardle.oncology.wisc.edu/sugden/Protocols/PDF files/Association of protein ... [阅读全文]
Mirmira Laboratory at the University of VirginiaThere are multiple variations to this protocol but we find that this one works well in all cases we tested.
http://faculty.virginia.edu/mirmira/resource ... [阅读全文]
Cepko/Tabin Lab Harvard University http://axon.med.harvard.edu/~cepko/protocol/mike/E1.html Cepko/Tabin Lab Harvard University http://axon.med.harvard.edu/~cepko/protocol/mike/E1.html Solutions
10X A ... [阅读全文]
John Mundy Institute of Molecular Biology Copenhagen Denmark http://www.arabidopsis.org/info/Protocols_Mundy2.jsp#Gelshift 1)Nuclear extract: see nucprep.ptc & nucext.ptc extracts should be at lea ... [阅读全文]
Caltech Genome Research LaboratoryCalifornia Institute of Technology http://www.tree.caltech.edu/protocols/agfp.html
Restriction digests consist of:
15.75 ml ddH2O
1 µl 10 X buffer B (Boehring ... [阅读全文]
Can anyone recommend a good kit for genome walking? I would like to find out the promoter sequence of a known gene. Is genome walking the way to do it?
thanks a million for any suggestions!
-kiwi-
Kiw ... [阅读全文]
genevasans-serif" size="2"Protocol written by Minoru Toyota
genevasans-serif" color="#932653" size="2"1. Materials
genevasans-serif" size="2"1.1. MCA
genevasans-serif" s ... [阅读全文]
DNaseI Footprintint
Solutions
10X Binding Buffer
200 mM Tris 8.0 200 m l 1M Tris pH 8.0
500 mM NaCl 100 m l 5M NaCl
10 mM EDTA 20 m l 0.5 M EDTA pH 8.0
680 m l Q
store at room temperature
DNaseI Dilut ... [阅读全文]
An end labeled DNA probe is incubated with a purified DNA-binding factor or with a protein extract. The unprotected DNA is then digested with DNase I such that on average every DNA molecule is c ... [阅读全文]
FINGERPRINTING PROTOCOL (AGAROSE GEL) *
Restriction digests consist of:
15.75 ml ddH2 O
1 µl 10 X buffer B (Boehringer Mannheim)
0.25 µl HindIII (40 U/µl)
3 µl DNA
Set up ... [阅读全文]