Kits RT-PCR
Conservation of STI samples for PCR
In general terms, the following is recommended for PCR analysis of STI samples: they can be stored at 2-8°C for up to 72 hours after collection, or at a temperature between -25°C and -15°C for longer periods of time.
E484K mutation (RTPCR010)
The RTPCR010 reference does not include the Spike E484K mutation for detecting SARS-CoV-2 variants, as it does not provide any additional discrimination of variants when HV69/70 and N501Y are already used. Additionally, including it would limit the ...
No amplification of N with amplification of E targets (RTPCR002)
In the RTPCR002 kit, the lack of amplification of the N target, despite the amplification of the E target, is due to a mutation in the viral RNA at the probe's target sequence. We recently identified several SARS-CoV-2 sequences with a single ...
Recommended nucleic acid extraction kits
RT-PCR format tubes
Both the LP and LPP formats use white "low profile" opaque tubes that allow fluorescence reading from the top angle, like the ones seen in the following images: Both formats are compatible with the LightCycler 480, however, a special frame is ...
Results interpretation in the Vaginal Panel Real Time PCR kit (RTPCR005) in VirCom
In attached files.
Mycobacteria and RTPCR016
Below is a list of the results of RTPCR016 after analyzing different species of mycobacteria: The regions amplified in RTPCR016 are IS1081 for MBTC, ITS region for MAC and MABSC and the 16S rRNA gene for NTM.
Duration of PCR
The duration of the PCR cycles depends on the heating/cooling ramping of each instruments, however in standard qPCR cyclers RTPCR001 takes about 1h 30 min and RTPCR002/RTPCR003 is 1h 45 min.
Colour compensation in LightCycler 480
The LightCycler 480 instrument requires color compensation for a 4-plex assay, as outlined in its manual. Color compensation can be applied using a file from a different assay that is using FAM, HEX, Texas Red (ROX), and Cy5. In case no color ...
PCR recommendations
1.- Use of separate hoods for handling samples and positive controls and the reconstitution and dispensing of the mix. 2.- Use of filter tips by trained personal. 3.- PCR instruments should be in a dedicated physically separated area from Pre-PCR ...
Differences between RTPCR001 and RTPCR002
RTPCR001 and RTPCR002 compatibility
Assigning variants with CT>36 RTPCR010
Generally, if any of the Ct values for the virus targets (FAM, HEX or Texas/ROX) is above 36, the result should not be considered for lineage assignment. If the N target is in the range of >36 Ct <40 the results should be “SARS CoV2 POSITIVE, LINEAGE ...