starblue SG maybe the primers themselves dun work for those species and those fragments. haiz... genetic stuff are so uncertain. one moment they work for something, then u change a species, it doesnt work anymore. sianzzz...
alexkusu SG i mean..maybe u didnt insert them into the wells deep enuf..so float on the top n screw up results ma
starblue SG /raises eyebrow. that's what the loading dye is for. besides, i'm getting too pro at loading gels now.
starblue SG primates. i have 9 species' dna with me now. trying to sequence their MCPH1 gene. currently working on exons 5, 9 and 13.
the Bear SG i won't even attempt to pretend i know anything... so, all i can do is hope that you will figure things out soon good luck sb
starblue SG erm, no, not a new species. as in let's say the primers for Exon 5 works for orang utan, then i use it for spider monkey, it may not work. or for howler monkey, the Exon 5 works, but die die also cannot get its Exon 9. liddat loh.
hisoka SG yeah i meant that its a new species you using the primers/exons on btw so you now everydya busy in uni arh?
starblue SG not a matter of figuring out things. it's more like trial and error. cos we're working with primers and enzymes, and they may just not respond cos the temperature differs by 1 degree, or cos the dna differs by 1 base. even the seniors in my lab also can only advice me on what other stuff i can try, but they also cannot predict what i will get. one lady even said that if i get too desperate (like how she was last time), i can try to do something utterly counter-intuitive. like the usual method is to lower the annealing temp by 2 degrees each time it fails. instead, maybe can increase by 1 degree. that sort of thing. lolz.. it just gets on your nerves when nothin works after 6 attempts.
