e-myco TM Mycoplasma PCR Detection Kit ( ver. 2.0, for 20 μlrxn]

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1 For research purposes only. Not for use in diagnostic procedures for clinical purposes. For N TRO US ONLY. e-myco TM R Detection Kit ( ver. 2.0, for 20 μlrxn] SO 9001/14001 ertified ompany at. No Tests DSRTON are common and serious contaminants of cell cultures. t has been shown that 30% to 87% of cell cultures are infected with mycoplasma. Many mycoplasma contaminations, particularly in continuous cell lines, grow slowly and do not destroy host cells but are still able to affect various parameters, leading to unreliable or false results. These effects include changes in metabolism, growth, viability, DN, RN, and protein synthesis, and morphology. Testing for mycoplasma is an essential quality tool to assure accurate research and reliable biotechnological products. The e-myco (ver.2.0) product is a set of primers designed to detect the presence of mycoplasma that might contaminate biological materials such as cultured cells. onventional techniques that are used to detect mycoplasma involve culturing samples on selective media, which needs at least 1 week to obtain results, whereas by performing R with this primer set, which is based on conserved 16S rrn, detection results are obtained in a few hours. ecause the presence of contaminant mycoplasma can be easily detected by only verifying the bands of amplified DN fragments using electrophoresis, there is no need to prepare probes that are labeled with radioisotope, etc. You can determine the species groups of mycoplasma by sequencing analysis using the sequencing primers suggested in this manual. Furthermore, if you want to know the detailed species, you may perform R and sequencing from your designed primers. The adopted 8-methoxypsoralen (8-MO) is used to extinguish the template activity of contaminated DNs. 8-MO is known to intercalate into double-stranded nucleic acids and form a covalent interstrand t crosslink after photo-activation ti ti by incidentid light at wavelength nm. n internal of this product was constructed to identify false negative results in each reaction. The internal was designed in such a way that the sample primer pair was used to amplify the internal and target DN, which were differentiated by size. ach tube of the e-myco R Detection Kit (ver.2.0) contains all the components for R except for template: i- StarTaq TM DN olymerase, dnts, 10x uffer, primers, 8-MO, and internal for partial gene amplifications. So, you can just add your templates and perform the R reaction. KT ONTNTS and STORG e-myco R Detection Kit (ver.2.0) 96 tubes DNase/RNase-free Distilled Water 1 vial (1 ml) Store at -20. The e-myco Detection R Kit (ver.2.0) is a novell vacuum-dried premix type. We guarantee storage for 12 months at -20. OMONNT R Reaction volume 20 μl reaction e-myco TM R Detection Kit (ver. 2.0) i-startaq TM DN olymerase 2.5U hemical Stabilizer 1x Loading uffer 1x dnts 250 mm each Tris-Hl (ph 8.3) 10 mm Kl 50 mm Mgl mm rimer Sets 10 pmol / each nternal ontrol 8-MO (dissolved in DMSO) HRTRSTS Dried under intron s instruction (atent ending) remix -Thise-Myco R Detection Kit(ver.2.0) contains all the components for the R reaction. You just add template DN or samples. road Species Detection - You can detect five common cell culture-infecting species of mycoplasma and also other various species of mycoplasma (See Technical Guide). Species Determination - You can determine the species of mycoplasma by sequencing the amplified R products. nternal ontrol - nternal embedded in the product prevents misjudgment that possibly arises from an erroneous R test. limination of arryover ontamination System - 8-MO solution prevents carryover contamination by R products. LTONS The kit is used for the detection of mycoplasma species that are most commonly encountered in cell culture, including,,, M. orale, and choleplasma laidlawii. Furthermore, this kit can detect other various species of mycoplasma (See Technical Guide). ROTOOL : ell boiling method 95 10min [ Overview of Detection ] Sample (culture) e- Myco TM R Detection Kit (ver.2.0) 1 tube : 20 μl D.W. (as negative ) 2 tube : 10 μl supernatant + 10ul D.W. ROTOOL : Genomic DN method i-genomic T DN xtraction Mini Kit (at.no ) R Reaction Detection on agarose gel U irradiation (10 min) Discard R product tube 1 tube : 20 μl D.W. (as negative ) 2 tube : 1 μl DN + 19ul D.W. info@intronbio.com -1- intron OTHNOLOGY

2 ROTOOL You can use this protocol just for detecting the contamination of mycoplasma. However, if you want to perform genotyping for the detailed determination of species, please purify the genomic DN of suspected -infected cells using our i-genomic T DN xtraction Mini Kit (at. No ). You may use simply this protocol or your other general boiling methods. [ THNL T ] 1. Use clean, disposable gloves when performing the assay and make sure that the work area is clean prior to starting the assay setup. 2. Keep your reagents and R mixture tubes on a cold block during reaction setup. 3. Use positive displacement pipettes. 4. The amplification and detection areas should be physically separated; i.e., do not use the same bench area to set up the R reactions and run your gels. [ UTONS ] DO NOT expose to U irradiation, which activates 8-MO, if you want to determine the detailed species of mycoplasma by DN sequencing analysis. f you want to do genotyping, excise the target band from the agarose gel, then isolate the DN fragment using a gel extraction kit.(eg. MG-spin TM garose Gel DN xttaction, intron, at.no 17181, MGquick-spin TM R & garose Gel DN xtraction Kit, intron, at.no 17281) ROTOOL : Using the oiling xtract Method 1. repare cell suspensions from the test cell culture in a 1.5 ml tube. Then count cell numbers by general counting methods. You need at least 5x10 4 cells per test. Note 1: Harvest adherent cells with trypsin-dt solution using standard techniques. ipette 1 ml of T-treated adherent cells. Generally, with suspension cells, such as K562, you need not treat with T solution. We recommend that you count the cells. You should prepare at least 5x10 4 cells per test (see Technical Guide, >50,000 cells are needed to complete this protocol). Note 2: Strong mycoplasma infections are detected in as little as 20~100 cells, while weak infections require cells over 50,000 cells. You can dilute the template according to the infection rates you suspect. We recommend that you perform the R reaction after preparing serial dilutions of the straight supernatant to obtain optimal results. 2. Transfer the counted cells (over 5x10 4 cells) to a 1.5 ml tube. Spin the tube in a microcentrifuge for 10~15 seconds. arefully decant the supernatant. 3. Resuspend the cells in 1 ml of sterile S or DS solution for washing. 4. Spin the tube in a microcentrifuge for 10~15 seconds. arefully decant the supernatant. Option : Repeat this wash step once more. 5. Resuspend the cell pellets in 100 μl of sterile S or DS solution. Note : f you want the best result, use of S solution is better than Tris (10 mm, ph 8.5), T (10 mm Tris, 0.1 mm DT), or autoclaved DW. 6. Heat the samples for 10 min, and vortex for 5-10 sec. Then, centrifuge for 2 min at 13,000 rpm with a tabletop centrifuge (at RT). 7. Transfer an aliquot of the heated supernatant to a fresh tube. This supernatant will be used as the template in the R. 8. dd 10 μl of the template to each tube of e-myco R Detection Kit (ver. 2.0), and then resuspend after adding 10μl of sterile water for a 20-μl R reaction volume. 9. erform R reaction as in the following table. Note : We recommend that you perform one negative reaction by adding 20μl of sterile water. 10. For analysis by electrophoresis, use 5 μl of each tube. 11. R products should be discarded after U irradiation (10 min) to prevent carry-over contamination. Note: ontamination of DN is a serious problem of R. lease discard R products after U irradiation (365 nm) to prevent carry-over contamination. ROTOOL : Using genomic DN as a template 1. dd purified genomic DN as a template using the i-genomic T DN xtraction Mini Kit (at.no ) to each tube of e-myco TM R Detection Kit (ver. 2.0), and then resuspend after adding sterile water for a 20-μl R reaction volume Note: ppropriate amounts of DN template sample: genomic DN, 50 ng 100 ng 2. Follow protocol from step 9. Note: We recommend that you perform one negative reaction by adding 20 μl of sterile water. TROULSSHOOTNG GUD 1. No band in positive sample heck nternal band : f internal band is seen, R has been performed properly; it is not a problem of the product. heck template quality :ven though DN is isolated from the sample, the R reaction can be inhibited depending on DN purity in some cases. n this case, extracted DN should be diluted 10 times with D.W. and used to perform R again. heck R machine : The problem can be caused by the R machine. lease check the temperature and make sure to check that the machine is working properly. 2. No internal band heck template concentration : ompetition can occur by high template concentration. lease proceed with a lower concentration of DN. heck template quality : ven though DN is isolated from the sample, the R reaction can be inhibited depending on DN purity in some cases. n this case, extracted DN should be diluted 10 times with D.W. and used to run the R reaction again. f still no band is seen, please inquire with our technical support staff. 3. mplicon bands in the negative heck contamination of D.W. : D.W. can be contaminated. erform R again with fresh sterile water. heck contamination of lab instruments and other environments : We recommend that you use filter tips to reduce contamination. We recommend that you use a pipette after sterilization. roceed with all procedures on a clean bench and keep the location where your procedures are performed sterile. 4. oor resolution on agarose gel We recommend using a 1.5~2% agarose gel. We recommend electrophoresis for 40 min at 100 /14 cm using a 6-cm long 2% agarose gel. 35 cycles R ondition Temp. Time nitial denaturation ti 94 o 1 min Denaturation 94 o 30 sec nnealing 60 o 20 sec xtension 72 o 1 min Final extension 72 o 5 min e-myco R Detection Kit ver. 2.0 T F

3 RNL OF MYOLSM DTTON THNL GUD SS DTRMNTON Y SQUNNG NLYSS The newly developed d e-myco M R Detection ti Kit (ver.2.0) is a highlyhl The sequence of amplified R products differs slightly from species to species. You sensitive R assay that detects various species that may contaminate can determine approximately the species by sequencing analysis with cell culture samples. The primer sets primarily allow for detection of major the following primers. lease refer to the phylogenetic table on the next page. For mycoplasma species in cell culture contaminations (,, M. more detailed species analysis, you should perform additional R reactions with fermentans,, ) as well as choleplasma laidlawii. Furthermore, your designed primers. you can detect various mycoplasma species with this kit (see below). t is a quick, simple, reliable, and cost-effective method for regularly monitoring cells for We list only the Forward primer sequences. lease synthesize the primer, and mycoplasma detection. then analyze by general sequencing methods. The primer sets used in the kit are derived from a highly conserved region within the Sequencing primer sequences : 16S rrn gene and can detect very low levels of contamination. The rrn gene Forward 5 - GT TG T TGG TG T-3 (20 mer) sequences of prokaryotes, including mycoplasma, are well conserved, whereas the Forward 5 - GT TG GT GT G-3 (20 mer) lengths and sequences of the spacer region in the rrn operon differ from species to species. So, you can determine the species by sequencing analysis. NLYT NFORMON Table 1 shows the broad species of mycoplasma detected by this kit. s shown, this kit can detect a broad range of mycoplasma with high specificity and sensitivity. The name comes from the Greek words mykes (fungus) and plasma (formed) and was proposed in the 1950s. is a genus of small bacteria that lack cell walls. Many species were purified and characterized from various origins such as cell culture, human, and cows. We classify them briefly by origin. Target rimers The target regions in this kit are divided into seven M types (M1~M7) and one type for detecting the bulk of the species in the genus. The designed primers are sufficient to detect major contaminants in cell cultures such as,,, M. hyorhinis,, and as well as other broad species of mycoplasma. R roduct The size of DN fragments that are amplified by the specific primers in this kit is about 270 bp. However, the sizes of R product differ slightly from species to species (268 bp~277 bp). You can confirm by sequencing analysis after T/ vector cloning and other cloning methods. ell culture Human vian L M N Guinea ig Squirrels Turkey D orcine O uma ovine anine F Ovine Q rimates G quine R Lion H Murine S Monkey nsect T Mink J Goat U Hamster K Geese guana Designed rimer * M1 Standard 15 T 16T 17 T 18T M6 18T, 20T M7 8T only * Not revealed primer sequences R 268 bp 269 bp 270 bp 271 bp 272 bp 277 bp Table 1. Species Detected by e-myco Kit (ver.2.0) [Note] The R primers used in this kit differ from the sequencing primers. We do not list the R primer sequences contained in this kit. RTL SQUNS OF MJOR ONTMNNTS N LL ULTUR The following sequences are partial sequences of major contaminants in general cell culture. You can classify the species by sequencing analysis. These sequences are the partial sequences of R products M. adleri M. agalactiae M. alkalescenns M. anseris M. arthritidis M. auris M. bovigenitalium M. bovirhinis M. bovis M. buccale M. californicum M. canadense M. caviae M. citelli M. cloacale / J F K / HF L M N rimer R M. columbinasale M. columbinum M. equirhinis M. falconis M. felifaucium M. gallinarum M. gateae M. hominis M. hyosynoviae M. iguanae M. indiense M. iners M. leopharyngis M. maculosum G O /D Q R rimer M1 M6 R M. meleagridis M. moatsii M. mustelae M. opalescens M. oxoniensis M. penetrans M. primatum M. pulmonis M. salivarium M. spermatophilum M. sualvi M. subdolum M. synoviae M. verecundum S T U Q H G rimer M7 R e-myco R Detection Kit ver T F

4 THNL GUD THNL NFORMTON This e-myco R Detection Kit (ver.2.0) will provide a sensitive means to detect mycoplasma contamination in cell lines. Under optimal conditions, templates derived from supernatants of an infected cell culture will yield a maximum signal in the R reaction, whereas an uninfectedcelllinewillyieldnorproducts. Undoubtedly, there will be variations in cell numbers, infection amount, and templates that may contribute to signal differences in your experiments. t is recommended that you use cultured cells that have cultivated for 3~6 days after subculturing as a sample for mycoplasma detection. You may not detect mycoplasma infection efficiently when you use cells that are not or shortly cultivated. The R amplification efficiency varies by mycoplasma infection range. Strong mycoplasma infections are detected in as little as 10~100 cell equivalents, while weak infections require cell equivalents from the 5000~50,000 range. So, we recommend that you plan various cell numbers in preparing R templates from the cultured cells by using the boiling method. lease refer to Fig. 2. f you perform genetic analysis for determining more detailed species, please extract the DN and apply it to the R process. We recommend that you use our i-genomic T DN xtraction Mini Kit (at. No ). HYLOGNT NLYSS TL The following phylogenetic analysis table shows the classification based on the sequence variations of R-amplified products. This cluster can be changed by which sequences are based on. This cluster is just a reference table. With a suggested sequencing primer, you can approximately determine the species. For example, because the cluster between and M. gallinarum is different, you can simply classify the species with just sequencing analysis. However, there is no difference between M. agalactiae, M. caviae,, and M. opalescens. n this case, you can t determine the detailed species with only this kit and a suggested sequencing primer. f you want to know the detailed species, you have to synthesize your own R primers, and then analyze by sequencing analysis. M. agalactiae M. caviae M. opalescens M. gallinarum M. adleri M. bovigenitalium M. californicum M. phocirhinis M. verecundum M. spermatophilum M. felifaucium M. maculosum M. leopharyngis M. meleagridis M. iners M. bovis M. columbinum M. columbinasale M. primatum M. lipofaciens M. citelli M. oxoniensis M. synoviae M. mustelae M. bovirhinis M. moatsii M. sualvi M. subdolum M. auris M. alkalescens M. anseris M. hominis M. cloacale M. buccale M. equirhinis M. canadense M. gateae M. hyosynoviae M. salivarium M. indiense M. falconis M. arthritidis M. iguanae M. pulmonis M. penetrans XRMNTL NFORMTON Detection limit K562 cell (-infected ) : small cell numbers, such as 12 cells K562 gdn (-infected ) : small quantities, such as 3.25 pg : small copy numbers, such as 20 cfu/ml 1) Result for the various concentration of template DN M nternal Fig.1. detection was performed for genomic DN Genomic DN was isolated from -infected K562 using the i-genomic T DN xtraction Mini Kit (17341). The isolated gdn was serially diluted for R of mycoplasma detection. These results show that it can be applied to mycoplasma detection with small quantities, such as 3.25 pg of gdn Lane M, 100bp DN Marker; lane, nternal,lane 1, 50 ng; lane 2, 25 ng; lane 3, 12.5 ng; lane 4, 6.3ng; lane 5, 3.2 ng; lane 6, 1.6 ng; lane 7, 800 pg; lane 8, 400 pg; lane 9, 200 pg; lane 10, 100 pg; lane 11, 50pg;lane 12, 25 pg; lane 13, 12.5 pg; lane 14, 6.3 pg; lane 15, 3.25 pg 2) Result for the various cell number M nternal Fig.2. detection was performed using the e-myco TM R Detection Kit (ver.2.0) method detection from cell lysates of -infected K562 using the e-myco TM Detection Kit (ver.2.0). The -infected K562 cells were serially diluted for R of mycoplasma detection and then R was performed per the e-myco TM Kit s protocol. These results show that it can be applied to the mycoplasma detection with small cell numbers, such as 12 cells Lane M, 100bp DN Marker; lane, nternal ; lane1, 2x10 5 ; lane 2, 1x10 5 ; lane 3, 5x10 4 ; lane 4, 2.5x10 4 ; lane 5, 1.25x10 4 ; lane 6, 6.25x10 3 ; lane 7, 3.125x10 3 ; lane 8, 1.56x10 3 ; lane 9, 7.8x10 2 ; lane 10, 3.9x10 2 ; lane 11, 1.9x10 2 ; lane 12, 96; lane 13, 48; lane 14, 24; lane 15, 12 3) Result for copy number M Fig.3. detection was performed using the e-myco TM R Detection Kit (ver.2.0) method detection from fermentans using the e-myco TM Detection Kit. were serially diluted for R of mycoplasma detection. These results show that it can be applied to mycoplasma detection with small copy numbers, such as 20 cfu/ml Lane M, 100bp DN Marker; lane1, 6.6x10 5 ; lane 2, 3.3x10 5 ; lane 3, 1.65x10 5 ; lane 4, 8.25x10 4 ; lane 5, 4.12x10 4 ; lane 6, 2.06x10 4 ; lane 7, 1.0x10 4 lane 8, 5.1x10 3 ; lane 9, 2.5x10 3 ; lane 10, 1.28x10 3 ; lane 11, 6.4x10 2 ; lane 12, 3.22x10 2 lane13, 1.61x10 2 ; lane 14, 80; lane 15, 40; lane 16, 20 limination of arryover ontamination M nternal 1) 1 st R nternal Fig. 4. detection was performed for genomic DN Lane M, 100bp DN Marker; lane 1, nternal ; lane 2, 25pg; lane 3, 12.5pg; lane 4, 6.3pg; lane 5, 3.25pg 2) 2 nd R M 1 w/o U w/ U Fig. 5. U irradiation (10min) of st R template Lane w/o U, without U irradiation; Lane w/u, with U irradiation (10 min); lane M, 100bp DN Marker ; lane 1, R product (1 μl) used from fiq. 4. lane 2; nternal lane 2, R product (1 μl) used from fiq. 4. lane 3; lane3,r product (1 μl) used from fiq.4. lane 4; lane 4, R product (1 μl) used from fiq. 4. lane 5; lane 5, R product (1 μl) used from fiq. 4. lane 2; lane 6, R product (1 μl) used from fiq. 4. lane 3; lane 7, R product (1 μl) used from fiq. 4. lane 4; lane 8, R product (1 μl) used from fiq. 4. Lane 5 e-myco R Detection Kit ver. 2.0 T F

5 THNL GUD RTL SQUNS OF MYOLSM SS MLFD Y e-myco Detection Kit (ver.2.0) > ii > M. columbinasale l > M. meleagridis GTTGGTGTGGTGTGGTGTGTTGGGGT GTTGGTGTGGGTGTGTTTGTGTGG GTTGTTGGTGTGTGTGTTTGTGTGG GTGTGGTTGTTGTTTGTGGTGTGTGGTT TTTGGGGTGTTTGTGTGGTGTGTTGG TTGTGGGGTGTTTGTGTGGTGTGTTGG GTGGTTGGGGGGGGTGGTTGTTGTTTT TGGTGTTGGTTTGTTTGGTTGGG TTTGGTGGTGTTTTTGTTTGGTT TTTGGTGGGTTTTTGTTTTGGTT >M.adleri GTTGTTGGTGTGTGTGTTTGTGTGGGG TTGGGGTGMTTTGTGTGGTGTGTTGG > M. agalactiae GTTGTTGGTGTGTGTGTTTGTGTGGGG > M. alkalescens TTTGGTGGGGTTTTTGTTTGGTT > M. anseris TTTGGTGGGTTTTTGTTTGGTTG > GTTGTTGGTGTGGTGTGTTTGTGGTGGGG TTTGGTGGGGTTTTTGTTTGTGTT > M. arthritidis GTTGTTGGTGTGGTGTGTTTGTGGTGGTT TTGGGTGTTTGTGTGGTGTTGTGG GTGTTGGTTGGGGGNGGGGTGGGTGTGGTTT TTTGNNTGGGGTTTTTGTTTGGTT >M.auris TTTGGTGGGGTTTTTGTTTGGTT > M. bovigenitalium GTTGTTGGTGTGTGTTGTTTGTGTGGGG TTTGGTTGGGGGGGGGTGGGTGTGGTTT TTTGGTGGTGTTTTTGTTTTGGTT > M. bovirhinis GTTGTTGGTGTGTGTGTGTGTTGTGTGGG GTTTGGGGTGTTTTGTGGTGTTGTGG GTGTTGGTTGGGGGTGGGGTGGGTGTGGTTT TTTGGTGGTGTTTTTGTTTTGGTT > M. bovis GTTGTTGGTGTGTGTGTTTGTTGTGGGG TTGGGTGTTTGTGTGGTGTGTTGG > M. buccale TTTGGTGGGTTTTTGTTTGTTTG > M. californicum GTTGTTGGTGTGTGTTGTTTGTGTGGGG TTTGGTTGGGGGGGGGTGGGTGTGGTTT TTTGTNNTGGTGTTTTTGTTTTGGTTG > M. canadense TTTGGTGGGGTTTTTGTTTGTGTT > M. caviae GTTGTTGGTGTGTGTGTTTGTGTGGGG > M. citelli GTTGTTGGTGTGGTGTGTGTTGTGTGG TGGGGTGTTTTGTGGTGTTGTGGGT > M. cloacale GTTGTTGGTGTGGTGTGTTTGTGGTGG TGTGGGTGTTTGTGTGGTGTTGTGGG TTTGGTGGTTTTTGTTTGGTTG > M. columbinum GTTGGTGTGGTGTGTTTGTGTGG TTTGGGGTGTTTGTGTGGTGTGTTGG > M. equirhinis GTTGGTGTGGGTGTGTTTGTGGTGGG TTTGGTGGTTTTTTGTTTTGTTG > M. falconis GTTGGTGTGGGTGTGTTTGTGGTGGYR RTTTGGGTGTTTGTGTGGTGTTGTGG GTGTTGGTTGGGGGGGGGTGGGTGTGGTTT TTTGGTGGGTTTTTGTTTTGGTT > GTTGGTGTGGGTGTGTTTGTGGTGGGG TGGGTGTTTGTGTGGTGTTGTGGGT GTTGGTTGGGGGGGGGTGGGTGTGGTTTT TTGGTGGGGTTTTTGTTTTGGTTG > M. felifaucium GTTGGTGTGGTGTGTTTGTGTGGG TTTGGGGTGTTTGTGTGGTGTGTTGG > GTTGGTGTGGTGTGTTTGTGTGGGG >M. gallinarum GTTGGTGTGGTGTGTTTGTGTGGGG > M. gateae GTTGGTGTGGGTGTGTTTGTGGTGGGG TTTGGTGGGGTTTTTGTTTGTGTT > M. hominis GTTGGTGTGGGTGTGTTTGTGGTGGG TTTGGTGGTTTTTGTTTGGTTG > GTTGGTGTGGGTGTGTTTGTTGGTGGT TTTTGGTGGTTTGTGTGGTGTTGTG GGTGTTGGTTGGGGGGGGTGGGTGTGGTT TTTTGGTGGTGTTTTTGTTTTGGT > M. hyosynoviae GTTGGTGTGGGTGTGTTTGTGGTGG TTTGGTTGGGGGGGGGTGGGTGTGGTTT TTTGGGTGGGGTTTTTGTTTGGTTG >M. iguanae GTTGGTGTGGGTGTGTTTGGGTGGT TTTGGGGTGGTTTGTGTGGTGTTGTGG GTGTTGGTTGGGGGGGGGGGGTGTGGTTT TTTGGTGGTGTTTTTGTTGTGTGTT > M. indiense GTTGGTGTGGTGTGTGTTTGTGGTGG TTGGGTGTTTGTGTGGTGTTGTGGG TTTGGTGGGGTTTTTGTTGGTTG >M.iners GTTGGTGTGGTGTGTTTGTGTGG TTTGGGGTGTTTGTGTGGTGTGTTGG > M. leopharyngis GTTGGTGTGGTGTGTTTGTGTGG TTGTGGGGTGTTTGTGTGGTGTGTTGG > M. maculosum GTTGGTGTGGTGTGTTTGTGTGGG TTGTGGGGTGTTTGTGTGGTGTGTTGG >M.moatsii GTTGTTGGTGTGGTGTGTTTGTGTGGG TTGTGGGTGTTTGTGTGGTGTTGTGG >M.mustelae GTTGTTGGTGTGTGTGTGTGTTGTGTGG TTGGGTGTTTTGTGGTGTTGTGGGT > M. opalescens GTTGTTGGTGTGTGTGTTTGTGTGGGG >M.orale GTTGTTGGTGTGTGTGTGTTTGTGGTGG TTGGGTGTTTGTGTGGTGTTGTGGG TTTGGTGGGGTTTTTGTTGGTTG > M. oxoniensis GTTGTTGGTGTGTGTGTGTGTTGTTGTGG TGGGTGTTTTGTGGTGTTGTGGGT > M. penetrans GTTGTTGGTGTGTGTGGTTTTTTGGTG GGTGTTGGTTGGTTTTTTGTGGGTGTTT GGTGTGGTTGGGGGGGTGGTGGGTG TTGTTTTGGTGTTTTGGGTTTGTTTG >M.primatum GTTGTTGGTGTGTGTGTTTGTTGTGGGG TTGGGTGTTTGTGTGGTGTGTTGG > M. pulmonis GTTGTTGGTGTGGTGTGTTTGTGGTGGTT TTTTGGGTGTTTGTTGGTGTTGTGG GTGTTGGTTGGGGGGGGGTGGGTGTGGTTT TTTGGTGGTGTTTTTGTTTGGTT > M. salivarium GTTGTTGGTGTGGTGTGTTTGTGGGG TGTTGGGTGTTTGTGTGGTGTTGTGGG TTTGGTTGGGGGGGGGTGGGTGTGGTTT TTTGGTGGGGTTTTTGTTTGGTTG > M. spermatophilum GTTGTTGGTGTGTGTGTTTGTGTGGG TTTGGGGTGTTTGTGTGGTGTGTTGG TTTGGTGGTGTTTTTGTTTTGTGTTG >M.sualvi GTTGTTGGTGTGGTGTGTTTGTGTGGG TTGTGGGTGTTTGTGTGGTGTTGTGG TTTGTGTTGGTGTTTTTGTTTTGGTT > M. subdolum GTTGTTGGTGTGGTGTGTTTGTGGTGGG TTTGGTGGGGTTTTTGTTTGGTT > M. synoviae GTTGTTGGTGTGTGTGTGTGTGTTGTGG TGGGTGTTGTTGTGGTGTTGTGGGT > M. verecundum GTTGTTGGTGTGGTGTGTGTTGTGTGGG TGGGGTGTTTTGTGGTGTTGTGGGT For more information, lease contact us without hesitation. info@intronbio.com e-myco R Detection Kit ver. 2.0 T F