just rinse the filter with 50 ml water first!
Thierry
On 21 Jul 2010, at 17:48, Danny Fuller wrote:
> Some manufacturers of filters have wetting agents in their filters, a reasonable precaution would be to discard the first 50ml or so of liquid through the filter. I have to agree, however, that degree of autoclaving can be a significant variable in Dicty growth.
>
> Danny
>
>
>> We also use Formedium and see little variation in doubling times. We are picky with the autoclaving, though. If your medium is brown Dicty won't grow well ... does not like caramel ! To test if that's the culprit, make a batch of sterile-filtered, non-autoclaved medium. Often relieves the problems.
>> Cheers,
>>
>> Thierry
>>
>>
>> On 21 Jul 2010, at 17:00, Robert Insall wrote:
>>
>>> We were one of the labs that suggested vitamin supplements to counter precisely this problem. Cells would grow at different speeds depending on background and in particular density (cells grew better above a threshold density in the region of 10^6/ml). The vitamins (and also not autoclaving the sugar; we made a 20x supplement with both) made the problem completely go away. They also slightly improved the doubling time. They do, however, inhibit development somewhat.
>>>
>>> Of course, that doesn't mean it'll answer your problem. Also, we moved labs and now it isn't apparently necessary - we haven't had a recurrence, and don't use the vitamins any more.
>>>
>>> Have you raised the issue with formedium? They would want to know. But our formedium HL5 seems fine.
>>>
>>> Robert
>>>
>>> On 21 Jul 2010, at 15:43, Christian Hammann wrote:
>>>
>>>> Is this also observed in other labs?
>>>> What is the usual doubling time of AX2 in your hands?
>>>> How constant is that doubling time?
>>>> How many of you have tried to supplement the medium with vitamins, as stated in the technical comments on Dictybase, and how successful was that?
>>>
>>> --------------------------------------
>>> Professor Robert Insall
>>> CR-UK Beatson Institute for Cancer Research
>>> Switchback Road, Bearsden
>>> Glasgow G61 1BD, UK
>>>
>>> Tel. (0/44) 141 330 4005
>>> Web http://www.beatson.gla.ac.uk/robert_insall
>>> --------------------------------------
>>
>> --
>> ================================================
>> Dr. Thierry Soldati, Senior Lecturer
>> Department of Biochemistry
>> University of Geneva
>> 30 quai Ernest Ansermet, Sciences II
>> CH-1211-Genève-4, Switzerland
>>
>> Tel: +41-22-379-6496
>> Fax: +41-22-379-3499
>> email: [log in to unmask]
>> http://www.unige.ch/sciences/biochimie/Research/Soldati.html
>> ================================================
>
--
================================================
Dr. Thierry Soldati, Senior Lecturer
Department of Biochemistry
University of Geneva
30 quai Ernest Ansermet, Sciences II
CH-1211-Genève-4, Switzerland
Tel: +41-22-379-6496
Fax: +41-22-379-3499
email: [log in to unmask]
http://www.unige.ch/sciences/biochimie/Research/Soldati.html
================================================
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