|
|
|
|
Visual-PCRTM
Mycoplasma Detection Kit
|
|
|
|
Mycoplasma has long been recognized as common contaminations of cells
culture. Among the mycoplasma detection methods, PCR is the highly sensitive, specific and convenient method1.
However, all of the available PCR kits have until now been a long and laborious process.
|
|
Visual-PCRTM
Mycoplasma Detection Kit is a simple, quick and sensitive way to visually detect
mycoplasma. It bases on our proprietary Visual-PCR technology. The technology makes PCR amplification to change color. For the very first time, PCR is capable of being visually monitored by naked eyes in visible light. DNA extraction is not required and crude sample is directly used as PCR template.
Post-PCR gel electrophoresis is unnecessary. Benefited from the
Visual-PCR technology, routine mycoplasma screening for cell
culture becomes as simple as pipetting sample.
|
|
|
|
|
Mycoplasma
Facts:
15%-80% cell cultures are
contaminated...
|
|
How to minimize
contamination
|
|
Challenge of
Mycopolasma
detection
|
|
|
PLEASE
CLICK HERE FOR DETAILS
|
|
|
Description:
|
|
|
Visual-PCRTM
technology is a major breakthrough of PCR. PCR reaction per
se generates sharp color-change which is easily detected by
naked eyes. PCR results, for the very first time, can be
visually read out without any machine or equipment. Prior-PCR DNA extraction/sample preparation and
post-PCR gel electrophoresis are not needed anymore.
Optimized primer sets covering all 6 common mycoplasma
species are chosen. PCR sensitive color-change component is
added into PCR cocktail. After PCR, positive reaction shows
a blue color, which indicates the presence of mycoplasma
DNA. Negative PCR reaction shows a purple color. The sharp
color change is easily identified by naked eyes (Figure
1,2,3,4). A variety of crude samples (cell cultures, frozen
cell lines, medium, serum, throat swab, etc) can be directly
used as PCR templates without DNA extraction and
pretreatment (centrifuge, boiling). Visual-PCRTM
kit represents the simplest PCR based assay for mycoplasma
detection.
|
| Advantages: |
- Revolutionary technology: visually
monitor PCR by naked eyes
- Sharp color change and can be easily read by naked eye (Figure 1,2,3,4)
- Hands-on time
less than 5 minutes
- Samples-to-results in 2 hours
- Crude samples
are directly used as PCR templates (Figure 3,4)
- No DNA extraction, No sample pretreatment (such as spin, boiling)
- No
post-PCR gel electrophoresis
- No
post-PCR pipetting
- Suitable for variety of crude samples: adherent or non-adherent cell cultures, frozen cell lines,
DMEM, RPMI-1640, serum, freezing solution, Penicillin/Streptomycin solution, non-essential amino acid, 1x PBS, throat-swab, etc. (Figure 3, 4)
- Demonstrated sensitivity: 10 copies/reaction (Figure 2)
- Detect numerous mycoplasma species/strains
(please click here for the list )
|
| Protocol: |
|
1.
Mix component A, B and C to prepare
complete-PCR-cocktail, aliquot 24ul per reaction.
2. Add 1ul crude samples and perform PCR. Visually check the
color of PCR tubes.
Fig1.
Typical color change of Visual-PCR
Mycoplasma Detection Kit.
PCR
reactions with or without template show
different colors. Positive is blue, and
negative is purple. |
|
|
Fig2.
Visual-PCR Mycoplasma Detection kit is
sensitive.
Different
copies of synthesized 16s RNA genes of
mycoplasma Orale were spiked into PCR
cocktail, and then PCR was performed. |
|
|
|
Fig3.
Crude samples are directly used as PCR
template.
Culture reagents (DMEM, FBS, RPMI-1640) were
spiked with mycoplasma (1000 copies/ul), and
then were directly used as PCR template. PCR
was then performed. |
|
Fig4.
Mycoplasma testing of a variety of crude
samples from a suspicious lab.
A
variety of samples from a suspicious
laboratory were tested for mycoplasma
contamination by Visual-PCR Mycoplasma
Detection Kit. Crude samples were directly
used as PCR templates without DNA extraction
or sample pretreatment. |
|
|
| Kit contents: |
|
Component A, B and C: to prepare
complete-PCR-cocktail
Component D: positive control DNA template (synthesized 16s RNA gene of Mycoplasma
orale)
|
| Storage: |
|
Product shipped on ice, store at -20°C upon arrival.
Complete-PCR-cocktail once prepared should be stored at 4°C. It is stable for 1 month.
|
| When To Test Mycoplasma: |
Example applications: |
|
•
Fail to reproduce published data
• Inconsistent, biased and skewed data
• Someone coughs or sneezes in the lab
• Culture a new cell line
• Obtain new cell lines from outside source
• Have not tested mycoplasma for 3 months
• Shared hood or CO2 incubator with others
• Cell culture manufacturing
• Production of cell related materials/products
|
|
•
Research cell cultures, frozen cell lines
• Cell culture media and supplements
• Serum
• High titer CHO cultures from bioreactors
• High titer NS0 cultures from bioreactors
• Cell culture vaccine manufacturing harvest
• Cell culture antibody manufacturing harvest
• Bioassay cell lines
• Stem cell cultures
• Lymphocyte proliferation cultures
• Cell and tissue therapy cultures
• Throat swab
• Surface wipes
|
| Technical references: |
|
1.
Young L, Sung J, Stacey G, Masters JR. Detection of Mycoplasma in cell cultures. Nat
Protoc. 2010;5(5):929-34.
|
|
|
|
|
|
| |
|
|
|
Ordering
Information |
|
|
| Cat# |
Description |
Price |
Doc |
Order |
|
|
| Request an Introductory Discount Code |
| If you are interested in testing the Visual-PCR Mycoplasma Detection Kit, please send email to ordering@gmbiosciences.com
to request your first kit at a 30% discount. The discount code is only valid for Cat# GM7036(15 tests). |
|
|
|
|
| |
|
|
|
