light sds bands

[vishwanathiari]

I am unable to get good bands in sds page i use 5% stacking gel and 12 % speparting gel, sample was denatured for 10 min at 90 oC satined in comassin blue for 30 min and destained overnight i have attached phot of gel 1st and last lane are markers lane 1,2,3,5,6,7 are protein samples 20 micro liters was loaded in biorad mini conatining 4x sample buffer diluted there is continues blue background in the lane i dont know why its is happening i ran at 60 volts constant for 3hrs aprox
please help me thank you
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[skumarmadhu]

make sure that the sample is in direct contact with the heat
heat samples for only five min, allow them to attain RT befor loading
make sure the conc of SDS

[vishwanathiari]

no now i got it right there was problem with sample preparation many lipds/ salts were interfering with the sample . i got kit from g bioscience page perfect then my bands are excellent