"Frontmatter". In: Plant Genomics and Proteomics

F
ULL
-L
ENGTH
cDNA S
EQUENCING
The approach to obtaining the sequence of a full-length cDNA clone is dif-
ferent from that used to generate EST data. Many of the full-length cDNAs
F
U L L
- L
E N G T H C
D N A S
E Q U E N C I N G
7 5
Full-length cDNAs
Protein function
Expression in heterologous systems
Use in transgenic analysis
Protein structure determination
using X-ray crystallography,
NMR, etc.
Genome annotation
including splice sites
using full-length
cDNA sequence
Full-length cDNA arrays
Expression profiling
FIGURE 4.2.
The contributions of full-length cDNA sequences to functional
genomics.

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are likely to be longer than the reads resulting from sequencing both the 3¢
and 5¢ ends of the insert. Therefore, additional sequencing strategies are nec-
essary to obtain the full-length cDNA sequence. The sequence must be of
high quality to identify possible SNPs and the transcripts from different
members of a gene family. The availability of such sequences is also impor-
tant for the confirmation of genes identified through computational genome
7 6
4. G
E N E
D
I S C O V E R Y
AAAAAAA AAAAAA
AAAAAAA
XTTTTTTTTTT
RE site primer adapter
AAAAAA
3'
5'
CAP
Full length cDNA
CAP
Biotinylation and First strand cDNA synthesis
RNase I treatment
Capture biotin-labeled CAP with Streptavidin
Hydrolyse RNA strand
dG tail 3' end of first strand cDNA
Second strand synthesis using oligoC+ attached RE site
Clone cDNAs using introduced RE sites
B
B
AAAAAAA
XTTTTTTTTTT
RE site primer adapter
AAAAAA