Multiprobe RNAse protection assay with internally labeled radioactive probes, generated by RT-PCR and NESTED PCR
نویسندگان
چکیده
منابع مشابه
Multiprobe RNase protection assay with internally labeled radioactive probes, generated by RT-PCR and nested PCR.
RNase Protection Assay (RPAs) is a highly sensitive and reproducible method of quantitating the levels of specific mRNA transcripts. The introduction of the commercially available Multiprobe RPAs allow comparing and quantifying the expression of up to different mRNA species in a single sample of 1-20 micrograms of total RNA. To generate probes which are not commercially available, we prepared h...
متن کاملDual-label detection of amplified products in quantitative RT-PCR assay using lanthanide-labeled probes.
Quantitative RT-PCR (QRT-PCR) enables the sensitive and specific detection of mRNA with a small copy number. We used the QRT-PCR method and dual-label analysis of amplification products for the detection of prostate-specific antigen (PSA) mRNA. The QRT-PCR assay employed a PSA-like internal standard (IS) mRNA, which was used to quantify the PSA mRNA copies and to control the variations during t...
متن کاملQuantitative RT-PCR using a PCR-generated competitive internal standard.
One approach to quantitate RNA is the use of a known number of competitive internal standard molecules that are reverse-transcribed and subsequently co-amplified under the same reaction conditions as the target sequence (3). This technique, called competitive reverse transcription polymerase chain reaction (cRT-PCR) has been applied to monitor the level of hepatitis C virus (HCV) RNA in patient...
متن کاملNested-RT-PCR and multiplex RT-PCR for diagnosis of rhinovirus infection in clinical samples.
Human rhinoviruses (HRVs) are positive-stranded RNA viruses belonging to the Enterovirus genus in the family of Picornaviridae. Identification of the specific strain in HRV disease has been difficult because the traditional serological method is insensitive, labor intensive, and cumbersome. With the fast progress in molecular biological technique, more sensitive and faster molecular methods hav...
متن کاملIdentification of spring viraemia of carp virus (SVCV) by combined RT-PCR and nested PCR.
A combination of single-tube reverse transcription (RT)-PCR and nested PCR was used to identify spring viraemia of carp rhabdovirus (SVCV) in infected cell cultures and fish tissues. Two pairs of specific primers (external and internal) were selected from the glycoprotein gene sequence. A specific product of 470 bp was amplified from RNA derived from 34 SVCV isolates including the reference str...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
ژورنال
عنوان ژورنال: Frontiers in Bioscience
سال: 1999
ISSN: 1093-9946,1093-4715
DOI: 10.2741/wolff