Search found 8 matches

by hara
Wed Feb 07, 2007 6:35 pm
Forum: Molecular Biology
Topic: PCR with only one primer
Replies: 17
Views: 30321

i think that if you want to seperate two pcr products with exactly the same size the next step after electrophoresis is to do a digestion of the pcr products with a restriction enzyme that cut the two sequences to different sites (so different fragments) and then do electrophoresis of the uncuts and...
by hara
Wed Feb 07, 2007 4:10 pm
Forum: Molecular Biology
Topic: decontamination of ethidium bromide
Replies: 6
Views: 4426

decontamination of ethidium bromide

is there an easy way that doesnt cost a lot to decontaminate an area around the electrophoresis tanks and the kodak imager from ethidium bromide? is it correct to do it someone with bleach? i need a solution that i can spread it to a large area and leave it for some hour and inactivate the etbr...th...
by hara
Wed Feb 07, 2007 3:56 pm
Forum: Molecular Biology
Topic: PCR primers
Replies: 2
Views: 1422

go to google then pubmed then entrez pubmed then nucleotide then blast then nucleotide-nucleotide(blastn) and then you put the sequence of the primer....try it and tell me if you suceed or else i help you...
by hara
Wed Jan 17, 2007 1:56 pm
Forum: Molecular Biology
Topic: loading buffer
Replies: 2
Views: 1895

loading buffer

why should i have a loading buffer that contains sds?
what is the difference with a loading buffer that contains just xylene cyanol FF,bromophenol blue, glycerol, EDTA,ddH2O?
if someone knows exactly the concetrations of the previous recipe i would like to let me know..thanks
by hara
Wed Jan 17, 2007 1:51 pm
Forum: Molecular Biology
Topic: agarose gel
Replies: 7
Views: 6843

agarose gel

i usually make my agarose gel this way: boil agarose powder in TBE 0.5X in a microwave oven then i let it cool a little and put ethidium bromide stir well and put it in the track to polymerize... recently i heard that some people put ethidium bromide in the TBE solution before making the gel has any...
by hara
Fri Jan 12, 2007 12:33 pm
Forum: Molecular Biology
Topic: dna storage
Replies: 10
Views: 5616

when you say tris pH=8.0 you mean to dissolve tris base in distilled water and then adjust the pH to 8.0 with HCl and sterilize by autoclaving?is this the solution you mean?thank you
by hara
Wed Jan 10, 2007 9:22 am
Forum: Molecular Biology
Topic: dna storage
Replies: 10
Views: 5616

thank you..but i worry that the EDTA (in tris) will cause problems later to my pcr as it chelates the divalent cations of Mg+2...but when i use depc H2O i have a difficulty in diluting my dna as it is acidic and the dna is diluting in slightly alcalic pH...THANK YOU FOR THE REPLY
by hara
Tue Jan 09, 2007 2:13 pm
Forum: Molecular Biology
Topic: dna storage
Replies: 10
Views: 5616

dna storage

when we complete the isolation of dna from leukocytes and before we do spectrophotometry to check the ratio and the concetration of it is it o.k to dilute it in depc water even if the ph of it is acidic?