GeneSpin Srl
Via Friuli, 51
20135 Milano
P.IVA C.F. 04520270960
Laboratory
and sales office
c/o Fondazione Filarete
Viale Ortles, 22/4
20139 Milano
tel. +39 02 56660199
fax. +39 02 537250
Cell Culture and Transfection
2X MycoPCR Master Mix is a 2X premixed, ready-to-use solution (GeneSpin proprietary formulation) containing XtraTaq Pol, dNTPs, MgCl2, specific mycoplasma primers set and stabilizers optimized for PCR-based mycoplasma detection in cell cultures. 2X MycoPCR Master Mix contains a combination of tartrazine and xylene dyes that allow gel loading and electrophoresis of the sample directly from the PCR tube, without further manipulation.
The primer set allows detection of various mycoplasma species (e.g., M. fermentans, M. hyorhinis, M. arginini, M. orale, M. salivarium, M. hominis, M. pulmonis, M. arthritidis, M. bovis, M. pneumoniae, M. pirum and M. capricolum), as well as Acholeplasma and Spiroplasma species, with high sensitivity and specificity.
cat. no.
STS-MICOP 150
amount
6x250ul
note
6x25 assay kit
STS-MICOP 50
2X250ul
2x25 assay kit
STS-MICOP 25
250ul
25 assay kit
FOR RESEARCH USE ONLY
SHIPPING
Shipped in green ice.
STORAGE
Store at 4C°
SHELF LIFE
12 months
FORM
Liquid white
PROTOCOL
1. Transfer 100-200ul of cell culture supernatant into 1.5ml centrifuge tube. Incubate the supernatant at 95°C for 5 minutes.
2. Centrifuge at maximum speed for 5 minutes.
3.Use 2-5ul of the supernatant as PCR template.
IMPORTANT! Before harvesting the supernatant from the cell culture, cells should cover approximately 90% of the growth surface! The supernatant may cause PCR inhibition in excessively dense cell cultures (>90%).
4. For optimum separation we recommend using a 2% agarose gel with TAE or TBE buffer used for electrophoresis.
5. Load all PCR product volume (20ul) directly onto the gel and perform electrophoresis.
6. When the electrophoretic run is completed, lay the gel onto UV transilluminator to detect the expected band.
7. IF the test is POSITIVE, a DNA band of 750bp will appear.
component
PCR assay set-up:
2X Muco PCR Mix
template from step 3
water
test sample
ctrl sample
10ul
10ul
2-5ul
2ul
8-5ul
8ul
20ul reaction
Cycling conditions:
Spin down the tubes/plate briefly to remove bubbles and place them into the cycler.
denaturation
denaturation (1)
annealing (2)
extension
final elong.
final step
95°C
95°C
58°C
72°C
72°C
4°C
5min
60 sec
90 sec
90 sec
forever
5 min
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www.genespin.com
info@genespin.com
administration@pec.genespin.com
