n***w
发帖数: 2405 | 1 Hi, I was doing transformation and after plating the culture for overnight
and no colonies grew. And there are many bubbles underneath the agar. Does
anyone have an idea how it happened and how to fix it?
Thank you so much. |
m******5
发帖数: 1383 | 2 making a new batch of plates…… |
n***w
发帖数: 2405 | 3 That's what I am doing now.
But I need to have an idea so I can avoid this problem when I make the
plates if it is due to something I didn't pay attention to in that process...
Thanks!
【在 m******5 的大作中提到】
: making a new batch of plates……
|
L**********O
发帖数: 1761 | 4 Make sure the antibiotics is correct.
And you can use the burner to get rid of the bubbles...
【在 n***w 的大作中提到】
: Hi, I was doing transformation and after plating the culture for overnight
: and no colonies grew. And there are many bubbles underneath the agar. Does
: anyone have an idea how it happened and how to fix it?
: Thank you so much.
|
T**********t
发帖数: 1604 | 5 That's two questions indeed.
That no colonies grew might result from many things. Not the right
antibiotic or not the right amount, or transformation went wrong... You need
to include at least a positive control and negative control each time when
you do transformation in order to troubleshoot.
Air bubbles in the plate usually happen when the media was too cold when
pouring the plates. The LB-Agar solidified before the accidentally trapped
air bubble had the chance to escape. The viscosity of the media also
increases when the temperature drops so air bubbles are more easy to be form
and trapped. |
n***w
发帖数: 2405 | 6 Thank you.
But I cannot see visible bubbles when I took the plates from 4 degrees.
After incubating at 37 degrees for
some time, they started to appear.
I will double check the antibiotic. Thank you.
need
when
form
【在 T**********t 的大作中提到】
: That's two questions indeed.
: That no colonies grew might result from many things. Not the right
: antibiotic or not the right amount, or transformation went wrong... You need
: to include at least a positive control and negative control each time when
: you do transformation in order to troubleshoot.
: Air bubbles in the plate usually happen when the media was too cold when
: pouring the plates. The LB-Agar solidified before the accidentally trapped
: air bubble had the chance to escape. The viscosity of the media also
: increases when the temperature drops so air bubbles are more easy to be form
: and trapped.
|
I*****h
发帖数: 470 | 7 大家都没回答到点子上，或者看出人家真正的问题
英文都挺溜啊
平板倒好了之后在室温放1-2天再放到4度
【在 n***w 的大作中提到】
: Thank you.
: But I cannot see visible bubbles when I took the plates from 4 degrees.
: After incubating at 37 degrees for
: some time, they started to appear.
: I will double check the antibiotic. Thank you.
:
: need
: when
: form
|
T**********t
发帖数: 1604 | 8 哈哈，英文问题我就用英文回答，中文问题就用中文回答了。。。
我实验室太脏，倒完平板之后我都是一凝好就放4度的，顶多过夜。在室温放1-2天万万
不敢，倒是没出现过lz说的那种一开始看不到气泡，incubate一阵之后出现气泡的情况。
【在 I*****h 的大作中提到】
: 大家都没回答到点子上，或者看出人家真正的问题
: 英文都挺溜啊
: 平板倒好了之后在室温放1-2天再放到4度
|
w**t
发帖数: 52 | 9 平板使用前在常温或者30度放1－2小时，等平板的温度升上来了再放到37度就不会有气
泡了。不过这些气泡不会影响你的转化结果，没有colony的原因很多。 |
n***w
发帖数: 2405 | 10 非常感谢！！
【在 w**t 的大作中提到】
: 平板使用前在常温或者30度放1－2小时，等平板的温度升上来了再放到37度就不会有气
: 泡了。不过这些气泡不会影响你的转化结果，没有colony的原因很多。
|
y***i
发帖数: 11639 | 11 铺板前冷却时要用stur bar搅动 （stur bar和LB一起 autoclave).
【在 n***w 的大作中提到】
: Hi, I was doing transformation and after plating the culture for overnight
: and no colonies grew. And there are many bubbles underneath the agar. Does
: anyone have an idea how it happened and how to fix it?
: Thank you so much.
|
