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支原体PCR检测试剂盒
英文名:Mycoplasma PCR Detection Kit
货号:8208
价格:¥7087.00
浏览量:1286次
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支原体PCR检测试剂盒描述:


       Mycoplasma, the smallest and self-replicating prokaryotes, are common contaminant of primary and continuous cell line cultures. It has been shown that micoplasma contamination affects cell growth, morphology and metabolism. Because of their small size and flexibility, mycoplasma can pass through commonly used filters of 0.2 μm. Furthermore, unlike bacterial and fungi, micoplasma contamination is resistant to commonly used antibiotics, and not easy to be spotted visually. These drawbacks emphasize the need for regular screening in control of mycoplasma contamination. Conventional screening methods include microbiological culture, fluorescent DNA staining and biochemical detection methods. However, mycoplasma culture is restricted to specialized laboratories and takes 2-4 weeks. Results of DNA fluorochrome staining are difficult to interpret due to the presence of broken nuclei. Various biochemical detections are time consuming and often lack of sensitivity. Recently, polymerase chain reaction (PCR) based methods have been developed as a quick and convenient detection of low level mycoplasma infection, which offers great advantage over the conventional methods because of its extreme sensitivity and specificity.


       Computer alignment studies of mycoplasmal 16S rRNA sequences have revealed the existence of highly conserved sequences. Primers designed according to these sequences allow the specific amplification of mycoplasma DNA fragment and thus highly sensitive detection of mycoplasma. ScienCell™ Mycoplasma PCR Detection Kit is a 16S rRNA-based PCR assay including 2X reaction buffer, primer set, as well as internal amplification control (IAC) and positive sample control. The genus-specific primer set we select recognizes the five typical contaminating mycoplasma species (i.e. M. hyorhinis, M. arginini, M. orale, M. fermentans, and A. laidlawi), which account for 98% of contaminations, as well as members of the genera Ureaplasma, Spiroplasma, and Acholeplasma, which account for the remaining 2% of contaminations. Eukaryotic and bacterial DNA is not amplified by this kit. The IAC is designed to be co-amplified simultaneously with the target mycoplasma sequence by the same set of primers. The resulted control band, which is distinguished from the target band by a difference in molecular mass, indicates the occurrence of DNA amplification. Therefore, false negative results due to the presence of PCR inhibitors in some cell cultures can be identified. Our kit also provides a positive sample control, which is noninfectious genomic DNA (gDNA) of M. fermentans. On the other hand, a no template (i.e. DNA-free DI H2O) negative sample control can help to determine whether there is a false positive result due to carry-over contamination. Each experiment should include both positive and negative sample controls, as well as the IAC.


产品使用说明:仅供科研研究使用

货号

8208

产地

美国

缩写

MYCO

规格

100 tests

用途

科研

保存

-20°C

运输

干冰

No Publication available at this time
Question:
       Dear ScienCell, We are interested in your mycoplasma kit (#8208) for use with iPSC cells grown in mTeSR media. We have a few questions about using the kit with these cells/media: 1) Do you know if the mTeSR (or other factors in these culture conditions) could act as a possible PCR inhibitor? (We are wondering if we need to do the genomic gDNA preparation.) 2) Do you know what the expiration date is for the kit (i.e., how long it is good for)? The cost for 100 tests is reasonable, but we want to make sure we use all 100 tests before the kit expires. In your reply, please also copy anna.l2.nelson@ucdenver.edu. Thank you very much for your time and assistance! Teisha Rowland


Answers:

       Posted by Manoj Sharma on Wednesday, January 27, 2016


Hi Teisha,

       1.) mTeSR should not have PCR inhibitors.

       2.) The shelf life of the kit is 3 months since one of the components is a long linear DNA which is not very stable during freeze/thaw.
       Thank you for your interest in ScienCell Products.
       Best
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