STRATAGENE MycoSensor PCR Assay Kit for Fast Mycoplasma Detection Manual
Contents
1. results due to carry over PCR product contamination The MycoSensor PCR assay kit provides a fast convenient method for the routine testing of cell cultures for mycoplasma and acholeplasma contamination The kit delivers consistent results and includes controls designed to eliminate false negatives by ensuring that the PCR amplification was successful and false positives due to reampli fication of the PCR amplicon are prevented when using the UDG decontamination treatment Contributing scientists Strat an Agilent Technologi 2008 Stratagene Dwight Dubois Scott Happe Marie Luna ee ee err nye Printed in USA April 2008 RE243 71109 5989 8260EN Patent pending See license reference 4 on page 102 www stratagene com Meet EGIES 16 3 812. DETECTS THE EIGHT MOST COMMON SPECIES RESULTS IN AS LITTLE eee Gel Based PCR Assay DETECTS AS FEW AS 10 COPIES OF MYCOPLASMA van for Fast Mycoplasma E COLI CONTAMINATION INCLONED TAQ IS NOT cree Detection The new MycoSensor PCR assay kit detects Mycoplasma contamination can interfere with your experiments on many levels This microor ganism can alter eukaryotic cell surface antigens chromosomal structure metabolic rates protein expression patterns and transfection efficiency Detection of mycoplasma is therefore critical for the reliability and reproducibility of experimental the fastest gel based PCR assay available The dara mycoplasma and acholeplasma infection in cell cultures in three and a half hours making it convenient kit allows you to routinely test for Mycoplasma Detection in Less Time the eight most common species of mycoplasma Stratagene was the first to develop a gel based PCR assay for mycoplasma detection Our new and acholeplasma improved kit the MycoSensor PCR assay kit features a reduced cycling time that allows you to detect mycoplasma contamination within three and a half hours including sample preparation Traditional mycoplasma detection methods such as om and gel electrophoresis Figure 1 culture testing staining and immunofluorescence detection are very time consuming insensitive and Teiran aeaea edetecis tae Giant difficult to interpret PCR amplific
3. ation provides a most common species of mycoplasma and acho quick and easy method for leplasma Table 1 mycoplasma detection E Reduce False Positives and False Negatives The kit includes a primer set two positive con trols an internal amplification control a dNTP dUTP mix and StrataClean resin Just add your choice of DNA polymerase we recommend Agarose gel analysis internal control Remove media or harvest cells Mycoplasma contamination internal control Treat with StrataClean Resin No Mycoplasma contamination Figure 1 The MycoSensor PCR Assay Kit Protocol STRATEGIES 16 3 STRATAGENE An Agilent Technologies Company Cell Biology Mycoplasma and acholeplasma species M pirum M salivarium M hominis A laidlawii M hyorhinis M arginini M fermentans M orale Table 1 Detect the Eight Most Common Species The MycoSensor PCR assay kit detects the eight most common species of mycoplasma and acholeplasma that contaminate cell cultures Taq The kit primers can detect infections from both supernatant and cell extract samples Cell Figure 2 canta inks f The MycoSensor PCR Assay Kit Eliminates False Negatives growth inhibiting or weak mycoplasma infections The kit detects the eight most common species of mycoplasma and acholeplasma can be detected by testing extracts made directly which account for gt 95 of mycoplasma contaminations in cell culture la
4. boratories While the kit has successfully detected mycoplasma from as few as 10 copies of from cells from as few as 10 copies of genomic mycoplasma DNA typical infections exceed 10 7 cfu ml of culture Lane 1 kb ladder DNA The primers are designed to prevent the Laine waters Ic otemplate contral amplification of E coli gDNA templates that some Lane 3 IC M orale Positive control 1 from the kit Lane 4 1IC A laidlawii Positive control 2 from the kit times contaminate recombinant Taq polymerase Lane 5 blank Lane 6 IC CHO cell sup Myco preparations Lane 7 IC HeLa cell sup Myco The two positive control templates validate that a i O a el polymerase mediated amplification has occurred MycoSensor PCR Assay Kit and confirm the size of the mycoplasma PCR product in the test samples If the test cell line is infected the mycoplasma primer mix yields a single 315 bp amplification product regardless of which species of mycoplasma is present Contents Catalog 100 assays 302108 The kit s internal control reduces false negatives by verifying that your PCR amplification was successful This template yields a second 500 bp band that is distinguishable from the mycoplasma target band Figure 2 The dNTP dUTP nucleotide mix contains dUTP for dUTP UNG decontamination treatment with uracil DNA glycosylase UDG not included with kit This eliminates false positive
Download Pdf Manuals
Related Search
STRATAGENE MycoSensor PCR Assay Kit for Fast Mycoplasma Detection Manual