To measure messenger RNA (mRNA) level of E2F5, total RNA was reversely transcribed using the Reverse Transcription System (Promega, Madison, WI).
Single-stranded complementary DNA (cDNA) was synthesized from 1 μg of total RNA using the Reverse transcription system (Promega, Leiden, The Netherlands) following the supplier's protocol.
For each individual sample, single-stranded complementary DNA (cDNA) was synthesized from 1 μg of total RNA using the Reverse transcription system (Promega, Leiden, The Netherlands) following the supplier's protocol.
Using the reverse transcription PCR, we evaluated expression levels of various antigen presentation-related genes, including LMP2, LMP7, MECL-1, PA28α, PA28β, TAP1, TAP2, and tapasin, in two oral squamous cell carcinoma cell lines, HSC5 and HSC7.
In this study, we identified a total of 32 alleles at five polymorphic SLA loci (SLA-1, SLA-3, SLA-2, DRB1 and DQB1) representing nine class I and seven class II haplotypes using the reverse transcription polymerase chain reaction (RT-PCR) sequence-based typing (SBT) method.
Approximately 2 ug of total RNAs were used for reverse transcription using the Reverse Transcription System (Promega, USA).
One microgram of RNA was used in the subsequent cDNA synthesis reaction, which was performed using the Reverse Transcription PCR kit (stratagene, USA).
cDNA was synthesized using the Reverse Transcription System kit (Promega) following manufacture's guidelines.
cDNA synthesis was performed using the Reverse Transcription System (Promega) according to manufacturers instructions.
Total RNA yield was determined by nanodrop quantitation and 1 µg was reverse transcribed using the Reverse Transcription System (Promega).
Samples were incubated at room temperature for 10 min; 42°C for 55 min; 95°C for five min using the Reverse Transcription System (Promega).
