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Asked 11th Nov, 2014
k Wright
ersity of Leeds
d protocol for the preparation of M9 Minimal Media?
are some M9 minimal media but previously the solution has become cloudy after preparation.

suggest what I am doing wrong?

Preparation Of Culture Media Cell Culture Techniques Bacterial Cell Culture Bacterial Cell Biology Microbiology Techniques

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salts solution (5X concentrate) :
View
of distilled/deionized water add: A minimal medium easiest to p

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n of (1 M MgSO4):
View
tilled/deionized water add:

SO4•7H2O

n of 40% glucose (w/v):

tilled/deionized water add:

se to stirring water in beaker; Do not attempt to add water to GLUCOSE!

n of (1M CaCl2):

tilled/deionized water add:

aCl2·2dH20

e with dH20

reparation of (1L of media) minimal medium:

9 salts solution

stilled water

agar media if agar plates are to be poured.

ing, swirl to mix evenly and "temper" at room temperature (until you can place your hand on the
ond) then add:

MgSO4 solution

CaCl2 solution

% glucose

dations
 Why is recombinant E.coli not
M9 minimal media ?
Question 37 answers
Asked 9th Sep, 2013
Priya Dharshini
I’m trying to grow recombinant E.
minimal media supplemented with
carbon source. I suspended 1 ml
(from LB media) in M9 media and
inoculate 5 mL of M9 media. But
growth in overnight culture.

The control strain(inoculated with

glycerol stock) has grown upto an
0.584 in overnight culture.
Powered by

This is my medium composition :

Na2HPO4 0.6%

KH2PO4 0.3%

NaCl 0.05%

NH4Cl 0.1%

CaCl2 0.1mM

MgSO4 1mM

ntial side effect to the pill: making MOPS 0.1M

women to read certain… Read More
Glucose 0.2%

My M9 medium pH is 6.88. How c

the medium so as to achieve goo
of the recombinant culture ? Or s
LB preculture for my later experim
cells adapt to m9 and grow

View

andro Martin 11th Nov, 2014

ter for Genetic Engineering and Biotechnology

ribe how you prepare the medium?

Wright 11th Nov, 2014

ersity of Leeds
 9 minimal media: I am making M9 minimal mediu
seeing no cell growth, where am
- autoclaved: 200 ml wrong?

%) - filtered: 20 ml Question 18 answers

Asked 5th May, 2012
- filtered and then autoclaved: 2 ml
Deleted profile
filtered then autoclaved: 0.1 ml My medium composition is

litre): Na2HPO4 0.6%

2O: 64g KH2PO4 0.3%

NaCl 2%

NH4Cl 0.1%

CaCl2 1mM

ave accidentally not pH'd to 7.4. Would this cause the precipitation? MgSO4 0.2%

dations I am autoclaving all medium cont

andro Martin except CaCl2(filter sterilized) and
11th Nov, 2014
ter for Genetic Engineering and Biotechnology (filter sterilized), I would be really
anyone can provide their thought
e nothing wrong with your recipe. Here we use half as much MgSO4, but I doubt that is the cause suggestion on this. I am using LB
es. You should not have to adjust pH, as that is taken care of by the combination of mono- and preculture, and when I inoculate i
hates. minimal medium, I used to give w
distilled water. But still my organis
ly optional. Can you leave it out and see if the same thing happens?
(Vibrio) unable to grow in this min
a Karimi Avargani 7th Jul, 2019
ersity of Isfahan
View
How can I calculate colony form
um (cfu) for bacteria??
salts solution (5X concentrate) : Question 67 answers

of distilled/deionized water add: Asked 1st Jan, 2016

Ozge Uysal
O4-7H2O
I want to calculate the colony form
bacterium which is frozen in glyce
4
will test an antibacterial surface s
know how many bacteria there ar
medium before putting them onto
Is there a way to calculate it?

r with dH20
View
utoclaving (or filter sterilization if autoclave is not available.) Why is bacterial density measu
600?
n of (1 M MgSO4):
Question 31 answers
tilled/deionized water add: Asked 2nd Feb, 2015
Raghvendra Ashok Bohara
SO4•7H2O
Why bacterial density is measure
n of 40% glucose (w/v):

tilled/deionized water add:

View

se to stirring water in beaker; Do not attempt to add water to GLUCOSE!

n of (1M CaCl2): Crystals precipitating out of M9

tilled/deionized water add: Question 11 answers

Asked 10th Oct, 2013
aCl2·2dH20
Maria C Davis
e with dH20 M9 medium prepared from M9 sa
from sigma. Medium is usually filt
reparation of (1L of media) minimal medium: followed by addition of glucose, C
MgSO4. Forms colourless crystal
9 salts solution 2-3 of storage at room temperatu
degrees. Can this medium be sto
stilled water
periods of time or does the mediu
agar media if agar plates are to be poured. freshly prepared?

ing, swirl to mix evenly and "temper" at room temperature (until you can place your hand on the
ond) then add:

MgSO4 solution View

How to exactly determine the O
CaCl2 solution E.coli suspension?
Question 54 answers
% glucose
Asked 6th Jun, 2013
dations
Peter Rehbein
ammad Yasir Malik 11th Nov, 2019 I am relatively new to the field of
national Islamic University, Islamabad
and could need some help of mor
experienced scientists in the field

I have grown an overnight-culture

e i need M9 media preparation exact protocol
BL21(DE3) in LB to an (apparent
how i can prepare
I tried to exactly determine the OD
the measured OD is by far above
uka Ranjan 2nd Feb, 2020 the suspension 1/16 and got an O
tre of Biomedical Research which calculates for a "real" OD6
= 6.24
to check the pH of your media
A dilution of 1/20 accordingly gav
0.17*20=3,4

Can you help by adding an answer? I understand that measuring OD6

dependent on the spectrometer a
and other things, but this differen
measurement undermines my co
our answer the acquired values... Is there som
a better control over the OD600
measurement? Which is the best
precise value of the OD of a high
bacterial suspension?

I then more or less "assumed" 6 t

correct OD600 of the overnight-cu
Add your answer gave me correct results after a se
up - a 1/60 dilution gave an OD o
tried to handle the overnight-cultu
solution" and pipetted the calcula
overnight-culture in the fresh incu
to get OD600 values of exactly 0.
starting OD600. I then measured
OD600, but these values were up
off the values I desired...
ns So my question is, is it possible to
create a suspension of e.g. OD60
from an overnight -culture of OD
y prepared culture media plates. A patient's life may depend upon your choice of culture would be the best way to do this?

Also, I want to try various media c

(minimal media), and inoculating
media with the LB-grown overnig
B Y Litsky would contaminate my prepared t
with small but unwanted amounts
only option here centrifugation an
ty of fluid culture media to small inocula resuspending the cells in the new
Which speed for centrifugation sh
so as to be able to separate the L
cells, but preventing the pellet fro
TTE · A FORGET too condensed, so that resuspens
possible?
dy of 7 freshly prepared culture media with the help of decimal dilutions of 21 pure cultures of
naerobic, shows that Trypticase Soy Broth (BBL) appears to be the best overall medium.
orites, such as VF and Robertson's, do not seem to fare as well under our experimental View
etween these, extremes can... Avoid calcium precipitation M9
medium?

dification to the Copacabana method for spreading transformation mixtures Question 4 answers
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Medium composition per liter:

Na2HPO4 6g

KH2PO4 3g

NaCl 0.5g

NH4Cl 1g

1 mL CaCl2 100mM

1 mL 1M MgSO4

3mL de Glycerol

Incubation conditions

25°C
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