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i) you don't need qPCR, just regular PCR with primers that would amplify the middle of the gene. Then sequence the PCR product and see if you have the truncated product or not. ii) Because you are studying a heritable cancer, it means that ALL the cells in the body inherit one bad mutation (the trun...
maybe that antibody is from a bad stock or something. You could try to borrow some from someone else and see if you get the same result. But really, if the longer blotting works, probably sticking with that is the wisest choice.
how long do you usually block for? Maybe 1 hour block is just not enough? Though I must confess, I've never seen an anomaly like yours, I don't understand why BSA would preferentially bind to where the lanes are and not in between
both actin and microtubules can grow both at the (+) and (-) ends. with actin, the reason actin in fact only grows at the plus end is that the concentration of monomers inside the cell is not high enough for the actin to grow at the (-) end also. If the concentration would be higher, actin would gro...
- Thu Apr 08, 2010 12:32 am
- Forum: Molecular Biology
- Topic: topoisomerases and gyrase
- Replies: 1
- Views: 10657
gyrase is a special kind of topoII that is present in bacteria to introduce negative supercoils into the DNA. Eukaryotes have nucleosomes to introduce the negative supercoils, so they don't need to have a girase.
- Sat Apr 03, 2010 9:01 pm
- Forum: Botany Discussion
- Topic: Why do intact chloroplasts fluoresce?
- Replies: 10
- Views: 11155
yeah mamoru you have it right! to put it simply, the electron capturing system that exists there isn't 100% efficient, not even close. Most of the time actually the energy just gets emitted as fluorescence. It's a good point - nothing in biology is ever 100% efficient, things are usually much below ...
- Sat Apr 03, 2010 2:45 pm
- Forum: Molecular Biology
- Topic: how making site-directed mutation on genome DNA
- Replies: 3
- Views: 2201
well first you have to figure out what your target sequence is (the sequence of the chromatin you want to mutate). Then make an artificial oligo that has the mutation, and introduce it into the genome by recombination insertion.
there is no way of estimating the error, you have to do the assay multiple times and calculate the standard deviation. It's a way of showing people that it wasn't just that one experiment that went well; doing it multiple times is the whole point
- Sat Apr 03, 2010 2:37 pm
- Forum: Cell Biology
- Topic: Tests to identify cell organelles?
- Replies: 6
- Views: 6224
well if you have organelles isolated and your procedure didn't **** them up, then you can do EM and just look at them, you should be able to recognize what they are if your fraction is pure enough. EM would also have the advantage that it will give you information on how pure and intact the organell...
the two tails create a unique shape of each molecule. If you only had one tail you could probably still form vesicles, but much smaller than those you are able to form with two tails, because a one-tail phospholipid would have much more of a conical shape