The SouthernBiotech Mycoplasma Detection Kit is designed to specifically detect potential ycoplasma contamination in cell cultures.The kit incorporates polymerase chain reaction (PCR) to amplify the conserved 16S ribosomal RNA coding region within the Mycoplasmagenome, thereby providing an extensive, highly sensitive, ａnd efficient detection method. Carefully determined primer sequences cover three genera of Mycoplasmatales (Mycoplasma, Acholeplasma, ａnd Ureaplasma) which allows the kit to detect over 95% of potential cell culture infections.

The PCR technology in the Mycoplasma Detection Kit is fast (results are typically obtained in less than 3 hours) ａnd easy to use. The kit is also highly sensitive ａnd can detect as little as 2-5 femtograms of Mycoplasma DNA in 100 µL of test sample supernatant. Eukaryotic ａnd bacterial DNA from cell  ulture supernatant is not amplified by the kit.

A sample cell line infected with Mycoplasma will generate a PCR product between ~448 bp to ~611 bp on an agarose gel, depending on the type of Mycoplasma present. A positive control (M. orale, 503 bp) is included to validate that the PCR amplification process has occurred as well as to confirm the size of the PCR product obtained in test samples. An internal control is also provided to eliminate

potential false negatives associated with PCR inhibitors.

Kit Components：

• User guide

• Primer set ａnd nucleotides (lyophilized) - Blue Cap

o Primer/dNTP mix (deoxynucleotide triphosphates including dATP, dCTP, dGTP, ａnd dUTP)

• Sterile PCR 10X reaction buffer (500 μL) - White Cap

o 100 mM Tris-HCl, pH 8.5

o 750 mM KCl

o 30 mM MgCl2

• Positive Control DNA (lyophilized) - Yellow Cap

o Non-infectious PCR product from M. orale

o Yields a 503 bp band

• Internal Control DNA (lyophilized) - Green Cap

o Non-infectious plasmid DNA including Mycoplasma-specific primer sequences

o Yields a 270 bp band

艾美捷支原体检测试剂盒中文说明：

组成：操作手册，设计引物/核苷酸Mix（dATP、dCTP、dGTP、dTTP），冻干粉，蓝盖装； 无菌PCR反应缓冲液10X，500uL，白盖装；阳性对照DNA（非传染性），冻干粉，黄盖装；内控对照质粒DNA（非传染性），冻干粉，绿盖装；试剂盒不含PCR仪，DNATag酶，PCR反应管，无DNA水

艾美捷支原体检测试剂盒特点：

1、试剂盒通过扩增支原体基因组中高度保守的16SrRNA编码区序列，实现培养细胞是否有支原体污染的检测

2、设计引物中含有支原体、无胆甾原体、尿素原体，能检测出95%以上的感染细胞；试剂盒操作简便，3小时内可出实验结果，灵敏性高，可以检测100uL样品上清液中2。5毫微微克的DNA，对于真核和细菌DNA不产生扩增效应

3、试剂盒提供的阳性对照在DNA电泳上会显示503bp的条带，提供的内控对照在DNA电泳上会显示270bp的条带（内控对照是PCR反应成功的标志）

相关文献：

1. Crittenden MR, Zebertavage L, Kramer G, Bambina S, Friedman D, Troesch V, et al. Tumor cure by radiation therapy ａnd checkpoint inhibitors depends on pre-existing immunity. Sci

Rep. 2018;8:7012.

2. Razidlo GL, Burton KM, McNiven MA. Interleukin-6 promotes pancreatic cancer cell migration by rapidly activating the small GTPase CDC42. J Biol Chem. 2018;293:11143-53.

3. Pishas KI, Drenberg CD, Taslim C, Theisen ER, Johnson KM, Saund RS, et al. Therapeutic targeting of KDM1A/LSD1 in Ewing sarcoma with SP-2509 engages the endoplasmic

reticulum stress response. Mol Cancer Ther. 2018;17:1902-16.

4. Bryant JD, Sweeney SR, Sentandreu E, Shin M, Ipas H, Xhemalce B, et al. Deletion of the neural tube defect-associated gene Mthfd1l disrupts one-carbon ａnd central energy

metabolism in mouse embryos. J Biol Chem. 2018;293:5821-33.

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