Who has positive controls for SARS-CoV-2 to test the following primers via Reverse Transcription Polymerase Chain Reaction (RT-PCR):
Proposed Annealing Temperature: 64.0°- 70.0°
Upper Primer:
MN988669.1:24180F23 (accession number:position direction length)
5' CAC TTC TGC ACT GTT AGC GGG TA 3'
Tm = 77.3° [%GC method]
Tm = 70° [2(A+T)° + 4(G+C)° method]
Lower Primer:
MN988669.1:24713R23
5' ATG AGG TGC TGA CTG AGG GAA GG 3'
Tm = 79.1° [%GC method]
Tm = 72° [2(A+T)° + 4(G+C)° method]
Amplified Product 556 bp (structural protein S of Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2))
Tm = 92.7° [%GC method]
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Upper Primer:
MN988669.1:26777F24
5' TGT CTT GTA GGC TTG ATG TGG CTC 3'
Tm = 77.3° [%GC method]
Tm = 72° [2(A+T)° + 4(G+C)° method]
Lower Primer:
MN988669.1:27010R25
5' AGC AAC AGT GAT TTC TTT AGG CAG G 3'
Tm = 75.7° [%GC method]
Tm = 72° [2(A+T)° + 4(G+C)° method]
Amplified Product 258 bp (structural protein M of Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2))
Tm = 95.1° [%GC method]
in silico PCR with coronavirus 2 isolate Wuhan-Hu-1, complete genome (NC_045512)
CDC provides the test kits for public health laboratories (PHLs) to perform real-time RT-polymerase chain reaction (rRT-PCR) detection of the SARS-CoV-2 virus (the virus that causes COVID-19) in respiratory specimens.
Hello Pavlos,
Asuragen has different Armored RNA controls which are stable, ribonuclease resistant RNA. You can reach them at [email protected] to learn more about what they can do.
Best of luck.
There are many laboratories around the world without any access to the free market and some of them cannot afford commercial kits.
In case of hight rate of corona-infections, they can detect the native virus by using the simple RT-PCR-Test with these primer-pairs.
If anybody needs help with the protocol or PCR-temperature-programm
can reach me at [email protected]
