How much kanamycin do you put on a LB plate?
Antibiotic Concentrations
| Antibiotic | Recommended Stock Concentration | Recommended Working Concentration |
|---|---|---|
| Coumermycin | 25 mg/mL (dissolve in DMSO) | 25 µg/mL |
| Gentamycin | 10 mg/mL | 10 µg/mL |
| Kanamycin | 50 mg/mL | 50 µg/mL |
| Spectinomycin | 50 mg/mL | 50 µg/mL |
What is the working concentration of kanamycin?
50 μg/ml
Working concentration is 50 μg/ml, or 25 μg/ml for low-copy plasmids like PACs. Stock solution is 35 mg/ml in water (kanamycin is insoluble in 50% alcohol).
How much antibiotic do I add to a pound of agar?
Preparation of LB-Ampicillin Agar To make 500 ml of LB agar containing 50 µg/ml ampicillin, add 2.5 ml of 10 mg/ml ampicillin stock. Ensure LB agar is cooled to 50°C before adding ampicillin, excessive heat will degrade the ampicillin.
Is LB Kan medium selective?
Application. Suitable for selective cultivation of E. LB Agar Kanamycin-50, Plates has been used in the screening of Agrobacterium tumefaciens and E. coli JM83 strain with pGD103 plasmid.
What is the difference between LB agar and LB broth?
Agar is a polysaccharide isolated from a plant and is used for microbial cultures on solid media. LB broth is generally used for inoculum preparation or start cultures in suspension medium. The only difference between broth and agar media is that broths do not contain an agar component… Thank you.
At what temperature does kanamycin degrade?
The results showed the reaction temperature had a significant impact on kanamycin degradation. The degradation half-life (t1/2) was shortened by 87.17-fold when the hydrothermal treatment temperature was increased from 100°C to 180°C.
How do you calculate antibiotic concentration?
Make sure units match!
- C1V1 = C2V.
- C = concentration; V = volume.
- Example: You have a stock solution of A at 100mg/ml. You want 150 ml at 100ug/ml.
- (100ug/ml)(150ml) = (100000ug/ml)(x)
- 15000 = 100000x.
- x = 0.15ml, or 150ul.
- Remember that you have to convert mg/ml to ug/ml, which you can do by multiplying by 1000.
How much volume of ampicillin stock 100 mg/ml will be added in 100 ml of LB broth so that final concentration would be 100 μg μl?
Ampicillin – The frozen stock solutions of ampicillin are at 50mg/ml and 100mg/ml in H2O, and are marked with a red sticker. The final concentration for LB liquid culture is 50ul/ml. To obtain this in 100ml (the amount in each LB bottle), add 100ul stock solution.
What type of medium is LB Kan?
LB Kan is Lysogeny Broth (LB) containing the antibiotic kanamycin. Because LB is a rich medium for growing bacteria, adding kanamycin provides a means of selecting transformants that have taken up plasmid DNA containing a gene which encodes resistance to kanamycin.
What does LB stand for in LB broth?
Lysogeny Broth
According to Bertani, LB has been variously misconstrued to stand for “Luria Broth”, “Luria-Bertani” medium, and “Lennox Broth”; however, the acronym originally stood for “Lysogeny Broth” (Bertani, 2004). The agar form of the medium should be designated LA but it is often referred to as LB.
Why is Luria broth perfect for E. coli?
Growth stops for lack of a utilizable carbon source. The carbon sources for E. coli in Luria-Bertani broth are catabolizable amino acids, not sugars. The widely used rich medium called Luria-Bertani broth is popular with bacteriologists because it permits fast growth and good growth yields for many species.
What happens if agar is too hot?
If the agar is too hot, the bacteria in the sample may be killed. If the agar is too cool, the medium may be lumpy once solidified.
What is the concentration of kanamycin in lb?
Kanamycin – The frozen stock solutions of kanamycin are at 50mg/ml in H 2O, and are marked with green. The final concentration for LB liquid culture for growing plasmids is 50ug/ml, and for cosmids is 20ug/ml. To obtain 50ng/ml in 100ml of LB, add 100ul stock solution, and to obtain 20ug/ml, add 40ul stock solution.
When do kanamycin plates start to lose sensitivity?
Plates start to lose sensitivity after 2 weeks; recommended Kanamycin plates only last a few weeks at 4°C; best to make fresh Kan plates every 2 – 4 weeks. Toxic to humans, use PPE! Rifampicin is TOXIC to humans, light-sensitive, and degrades in solution rather quickly.
Which is concentration of rifampin and kanamycin can I use to?
Usually we use rifampicin at 10mg/L and kanamycin at 50mg/L. I use to prepare Rif stock solution (10mg/mL) by dissolving it in methanol. As Rif gets precipitated during long storage, prepare the stock with minimum quantity.
How much Ampicillin is in lb liquid culture?
How to make LB liquid plus antibiotics: Ampicillin – The frozen stock solutions of ampicillin are at 50mg/ml and 100mg/ml in H 2 O, and are marked with a red sticker. The final concentration for LB liquid culture is 50ul/ml.