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

PDF

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

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

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www.genespin.com

info@genespin.com

administration@pec.genespin.com

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