1. S Vaibhav's Avatar
    My experiment was not successful. All the strains don't have the deletion I wanted to introduce. Man, that's ****ed up.
    What was the experiment about? : Curious:
    Chregu likes this.
    07-07-2014 11:02 AM
  2. sd173's Avatar
    The worst part is I can't move the same video that I downloaded through the WP 8.1 app since it's stuck in the app and not visible though the Files app.
    07-07-2014 11:03 AM
  3. Chregu's Avatar
    What was the experiment about? : Curious:
    I'm trying to create a bacteria strain that is very important for my experiments. It's already pretty sick and I need to knock-out an additional gene. It didn't work. It was the second time I tried...
    07-07-2014 11:09 AM
  4. Chregu's Avatar
    I have to try out the LG G3 in a store. I think it's an amazing phone that is much cheaper than the S5 or M8. But unfortunately it's also huge.
    07-07-2014 11:13 AM
  5. S Vaibhav's Avatar
    How do you create it(the strain)? Do you have to insert the desired gene into a vector and then it binds with the plasmid of the bacteria to produce the desired effect?
    07-07-2014 11:17 AM
  6. Chregu's Avatar
    How do you create it(the strain)? Do you have to insert the desired gene into a vector and then it binds with the plasmid of the bacteria to produce the desired effect?
    It's a double recombination with a suicide vector. To knock-out the gene first around 600 bp of the DNA upstream and downstream of the gene is cloned into a vector. After bringing the vector into the bacteria, these two regions then recombine with a very low efficiency and remove the gene while bringing in a antibiotic resistance cassette and a sacB, a special gene that is important later. By selection for the antibiotic resistance colonies with the recombination can by gained. These will then be grown in a liquid culture again to make a second recombination, losing the resistance cassette and sacB. You then do a second selection by growing the bacteria on sucrose. Sucrose kills all bacteria that have the sacB gene.

    The now yielded colonies have to be screened for lost antibiotic resistance (to avoid choosing bacteria with a mutation on sacB) and then a colony PCR is performed to confirm the knock-out.

    Everything worked until the last step. But no colony had lost the gene...
    sd173 and S Vaibhav like this.
    07-07-2014 11:24 AM
  7. sahib lopez's Avatar
    o.o that explanation to make a strain is complicated
    Chregu and sd173 like this.
    07-07-2014 11:29 AM
  8. Chregu's Avatar
    o.o that explanation to make a strain is complicated
    It's like with everything. It sound complicated but as soon as you do it on a daily basis it's just everyday work ;-)
    sd173 and S Vaibhav like this.
    07-07-2014 11:31 AM
  9. Chregu's Avatar
    It's raining heavily, no fun to cycle home. And then I still have to get groceries. Well, it's fine I guess.
    07-07-2014 11:36 AM
  10. sahib lopez's Avatar
    It's like with everything. It sound complicated but as soon as you do it on a daily basis it's just everyday work ;-)
    :P yeah let me get edumacated on micro organisms
    sd173 and Chregu like this.
    07-07-2014 11:38 AM
  11. sahib lopez's Avatar
    It's raining heavily, no fun to cycle home. And then I still have to get groceries. Well, it's fine I guess.
    I remember that my ex love to ride her bike in the rain >.<.... it might be fun :P for you
    07-07-2014 11:40 AM
  12. N_LaRUE's Avatar
    I remember that my ex love to ride her bike in the rain >.<.... it might be fun :P for you
    When it's a warm day and the rain is warm, riding your bike in it is fun. I did it many times when I was younger. We used to sing while riding ( very badly I might add).
    Chregu and kmji1957 like this.
    07-07-2014 11:57 AM
  13. sahib lopez's Avatar
    man the other forms here are about saying about the android powered lumia and having android apps running for the store. lets just ask BB how they are doing >.>
    sd173 likes this.
    07-07-2014 11:57 AM
  14. sahib lopez's Avatar
    When it's a warm day and the rain is warm, riding your bike in it is fun. I did it many times when I was younger. We used to sing while riding ( very badly I might add).
    I never did that because I never had a bike to ride in :( oh well but I did get to run in it :P . I do think its fun when its warm :3
    07-07-2014 11:58 AM
  15. N_LaRUE's Avatar
    I remember that my ex love to ride her bike in the rain >.<.... it might be fun :P for you
    Note. When meeting up with friend from for years ago keep talk of ex to minimum. Be nonchalant. Indifferent. That it's over and you moved on.
    Chregu likes this.
    07-07-2014 11:58 AM
  16. N_LaRUE's Avatar
    I never did that because I never had a bike to ride in :( oh well but I did get to run in it :P . I do think its fun when its warm :3
    I got my first bike at around 5 and my first two wheel around 8. After that I bought my own bike and paid for any mods myself. My last bike was a very heavy mountain bike. I haven't owned a bike since.
    kmji1957 likes this.
    07-07-2014 12:01 PM
  17. sahib lopez's Avatar
    Note. When meeting up with friend from for years ago keep talk of ex to minimum. Be nonchalant. Indifferent. That it's over and you moved on.
    lol she doesn't know :P and I wasn't planning to say anything unless she asks me XD . there was this one time when she did but I didn't tell her then :P
    07-07-2014 12:02 PM
  18. N_LaRUE's Avatar
    lol she doesn't know :P and I wasn't planning to say anything unless she asks me XD . there was this one time when she did but I didn't tell her then :P
    If it comes up just keep it short and simple. Some lady folk like to dig. So be wary.
    sahib lopez and Chregu like this.
    07-07-2014 12:03 PM
  19. sahib lopez's Avatar
    I got my first bike at around 5 and my first two wheel around 8. After that I bought my own bike and paid for any mods myself. My last bike was a very heavy mountain bike. I haven't owned a bike since.
    I did have a scooter ^.^ anyways I didn't know you knew about moding your bike o.o that's legit
    07-07-2014 12:04 PM
  20. sahib lopez's Avatar
    If it comes up just keep it short and simple. Some lady folk like to dig. So be wary.
    ok if you say so but I don't think I have to worry about it
    07-07-2014 12:05 PM
  21. N_LaRUE's Avatar
    ok if you say so but I don't think I have to worry about it
    That's fine. Enjoy your time with your friend. :)
    Chregu likes this.
    07-07-2014 12:07 PM
  22. N_LaRUE's Avatar
    I did have a scooter ^.^ anyways I didn't know you knew about moding your bike o.o that's legit
    I used to do all kinds of things. I know how to ice skate, skateboard, Rollerblade and ride a bike. Do some tricks here and there. A bit of fun. :)
    07-07-2014 12:08 PM
  23. sahib lopez's Avatar
    That's fine. Enjoy your time with your friend. :)
    thanks and I know I will >:)
    07-07-2014 12:10 PM
  24. sahib lopez's Avatar
    I used to do all kinds of things. I know how to ice skate, skateboard, Rollerblade and ride a bike. Do some tricks here and there. A bit of fun. :)
    o.o you were a street warrior ! I only know how to ice skate and ride a bike and that is about it . I would always fall of the skateboard and eat it by rollerblading.
    07-07-2014 12:12 PM
  25. MSFTisMIA's Avatar
    What a day so far. Everyone's pregnant and coworkers are cherry picking work. A great recipe for me to "go berserk" on some folks.
    07-07-2014 12:19 PM
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