Appeal No. 1996-2442
Application 08/062,021
approximately equal amplification of the different gene fragments). Page 238. On page
5 of the Examiner’s Answer, the examiner shows that the Tms of eight of the primers
(Gibbs’ first four primer pairs) range from 65.64°C to 76.55°C, based on primer length
and GC content, and some of the pairs have Tms within 5°C of each other.
Chamberlain discloses multiplex PCR using 1 :M each of twelve primers (six
primer sets), and 10 units of Taq DNA polymerase per 100 :l of reaction mixture.
There is no mention of primer Tms, but the reference teaches that combining multiple
primer sets in a single reaction requires modification of the annealing temperatures of
the individual primers, among other things. Page 11146.
Nedjar discloses co-amplification and detection of hepatitis C virus (HCV) and
human immunodeficiency virus (HIV) specific sequences in the same sample, using
nested primer pairs in the polymerase chain reaction (PCR). Nested PCR requires
sequential rounds of amplification, rather than simultaneous, because the sequence
amplified using the “inner” primer pair is embedded in the sequence amplified using the
“outer” primer pair.
Brytting discloses detection of conserved sequences from the immediate early
gene of human cytomegalovirus (hCMV) using nested primer pairs.
Findlay uses a “nucleic acid test article” with a specific nucleic acid probe
immobilized on its surface to capture complementary nucleic acid from a sample.
5
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