LEXSY Cultivation Kit T7-TR

Detailní popis

for inducible expression

For general laboratory use. Not intended for human or animal diagnostic or therapeutic uses.

Shipping: Shipped on dry ice/on gel packs

Storage Conditions: Store components as indicated on individual labels

Shelf Life: See individual components

Description:
The LEXSY Cultivation Kit T7-TR was developed for initial cultivation and establishment of Leishmania tarentolae laboratory strain T7-TR in suspension culture.

Content:
3 vials with 1.6 ml each of frozen glycerol stocks of LEXSY host T7-TR each
(Leishmania tarentolae strain T7-TR expressing bacteriphage T7 RNA
polymerase and TET repressor)
Upon arrival the glycerol stocks must be stored at -80 °C or inoculated into the provided LEXSY BHI liquid medium. Do not thaw and re-freeze the stocks.

50 ml LEXSY BHI liquid medium complete
For initial inoculation, contains Hemin, Nourseothricin (NTC), LEXSY Hygro and PenStrep.
Store at 4 °C, stable for 2 weeks

37 g LEXSY BHI powder
2x 18.5 g for preparation of 2x 500 ml LEXSY BHI growth medium.
Store at ambient temperature, stable for 3 years

2 ml Hemin stock solution, 500x
0,25 % solution of porcine Hemin in 30 % Triethanolamine for 1 L LEXSY BHI medium
Store at 4 °C in the dark, stable for 6 months

5 ml Pen-Strep stock solution, 200x
10.000 units/ml of penicillin (base) and 10.000 μg/ml of streptomycin (base) in 0.85 % saline, for 1 L LEXSY BHI medium.
Store at -20 °C, stable for 6 months

1 ml Nourseothricin (NTC) and 1 ml LEXSY Hygro
For maintenance of T7 polymerase and TET repressor genes.
Store at -20 °C, stable for 6 months

4 cryo vials with 0.4 ml 80% glycerol each, sterile
For preparation of glycerol stocks.
Store at ambient temperature, stable for 6 months

3 culture flasks for 10 ml medium
For initial inoculation.

Biosafety level:
1, Non-pathogenic

Source of wild type organism:
Tarentola annularis

Reactivation of LEXSY host:
Thaw glycerol stock on ice for ca. 20 minutes and inoculate the entire content of the vial into 10 ml of LEXSY BHI medium in one of the provided cell culture flasks. Incubate at 26 °C in the dark and dilute as required.

Preparation of LEXSY BHI growth medium:
Dissolve 37 g/l LEXSY BHI powder in deionized water and autoclave max. 15 min at 121 °C. Add Hemin, Nourseothricin (NTC), LEXSY Hygro and PenStrep to 1x final concentration. Store at 4 °C and use within two weeks.

Preparation of glycerol stocks:
Add 1.2 ml of growing culture (ca. 6x10⁷ cells/ml) to a vial with 0.4 ml 80 % glycerol, mix, incubate 10 min at RT, 1 h on ice and over night at -20 °C. Transfer to -80 °C. Strains can be stored this way for several years.

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