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准备工作： 细菌培养：小量提取质粒DNA 鉴定正确后，将菌液以1/50~1/20体积的比例接种到250ml液体培养基中，37℃振荡培养过夜至对数生长晚期。 注：培养体积与最终质粒DNA 的收获量并无线性关系。只要培养条件适宜，50ml的培养液有时也可得到总量高达1mg以上的质粒。 操作步骤 ： 1、细菌的收获：将菌液倒入合适的离心管中，8000g离心5分钟，弃上清，并在吸水纸上倒置离心管使上 ...

Hey everybody I am working on a side-directed mutagenesis with the Stratagene℃ quickchange mutagenesis xl-kit. Here is my problem: My plasmid is 17kb big. Until now I have had no colonies on my plate. ...

I am trying to extract DNA from mammalian blood but while doing the phenol extraction lot of Gelatinous material in aqeous phase creating problem in its separation. Can somebody tell me how to m ...

1.Grind in mortar and pestle or Waring blender with 5-7 volumes buffer A per g tissue. Use MCE at 350 l/L and if necessary with 5 ml 1 M DIECA/L. 2.Squeeze through cheesecloth two layers of Miracloth ...

Paula Marquardt & Craig Echt Published in: Echt CS May-Marquardt P Hseih M Zahorchak R. 1996. Characterization of microsatellite markers in eastern white pine. Genome 39:1102-1108 . comments can b ...

Purpose: Purification of oligonucleotides (which have already been purified by reverse phase) can increase the efficiency of site directed mutagenesis from around 30% to ~75% or greater in some cases ...

This is the URL to my gene of interest. I am trying to estimate the promoter region of this gene so that I can do Meth specific PCR on the promoter..the promoter region of this gene has not been posit ...

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 ...

This method is derived from a procedure developed by Toby Bradshaw and the Poplar Molecular Genetics Cooperative. We have tested the procedure with a variety of Populus species as well as tobacco and ...

QUOTE julianne: been using sod acetate- isoprop precipiation with no problems all the while. QUOTE Turtle: my suggestion would be to ethanol precipitate QUOTE for PCR product purification used for clo ...

I did stupid thing after cloning a sequence into a plasmid just find out not in frame with the following sequence. just need insert one single nucleoacid will be ok. Any method to insert only one nucl ...

Hi can anyone answer this I am having problems with my southern hybridization. I am wondering if the DNA transfers to my membrane. Is it possible that the DNA can wash into the solutions when denaturi ...

Materials: Silica Suspension: add 2 g of silica to 15 ml of H2O wash 3x by centifugation at 2000 x g for 2 min estimate vol of silica and resuspend in 2 vol H2O Silica Wash Solution: 50 mM NaCl 10 mM ...

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 ...

Caution: EtBr is a powerful mutagen and is moderately toxic. Wear gloves when working. When EtBr conc. is >0.5 mg/ml: 1.Add sufficient water to reduce the concentration of EtBr to <0.5 mg/ml. ...

I am working with very small amount human tissue. Is there anyone who had the experience of using glycogen with 100% ethanol in pheno/chloroform DNA extraction protocol ? Does it realy work to increas ...

Hancock Laboratory Methods. Department of Microbiology and Immunology University of British Columbia British Columbia Canada http://www.cmdr.ubc.ca/bobh/showmethod.php?methodid=67 METHOD: 1.Run gel a ...

Sample Preparation Cheek cells are obtained by rinsing the mouth with 25mls of any commercial mouth wash solution available for about 30sec the first thing in the morning. It is important not to brus ...

Hello group! I want to insert a small 30 bp sequence into a vector of mine using ds-oligo. I've created the oligos so that once annealed they have the correct overhangs of the restriction sites. As I' ...

Donis-Keller Lab Manual Department of Genetics in Washington University School of Medicine http://hg.wustl.edu/hdk_lab_manual/plasmid/plsmid08.html Purpose: To utilize competent E. coli bacteria to r ...