EP0544809B1
Expired / Lapsed
EP
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Methods of synthesizing oligonucleotides with random codons
Assignee: Applied Molecular Evolution Inc
•
Filing: 1991-08-20
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Granted: 1998-12-16
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Expires: 2011-08-20
Abstract
Abstract The invention provides a method of synthesizing oligonucleotides having random tuplets using individual monomers. The steps consist of: (1) sequentially coupling monomers on separate supports to form at least two different tuplets, the coupling is performed in separate reaction vessels; (2) mixing the supports from the reaction vessels; (3) dividing the mixed supports into two or more separate reaction vessels; and (4) repeating steps (1) through (3) one or more times in the reaction vessels of step (3), wherein the last step ends at step (2). Additionally, the oligonucleotides can be cleaved from the supports.
Inventors
- 👤 William D. Huse
Bibliographic Details
Application Number
—
Jurisdiction
EP
Data Source
google_patents
Last Synchronized
2026-11-04 17:25
CPC Classifications
C; C07; C07H; C07H21/00
Claims Analysis
Total Claims: 25
Claim 1
Independent Claim
84 words
A method of synthesizing oligonucleotides
having random tuplets using individual monomers comprising
the steps of: (1) sequentially coupling monomers on
separate supports to form at least two
different tuplets, said coupling being
performed in separate reaction vessels; (2) mixing the supports from the separate
reaction vessels; (3) dividing the mixed supports into two or
more separate reaction vessels; and (4) repeating steps (1) through (3) one or
more times in the reaction vessels of
step (3), wherein the last step ends at
step (2).
Claim 2
Dependent Claim
18 words
The method of claim 1 further comprising
step (5) cleaving said oligonucleotides with random tuplets
from said support.
Claim 3
Dependent Claim
11 words
The method of claim 1, wherein the tuplet is
a triplet.
Claim 4
Dependent Claim
18 words
The method of claim 1, wherein the number of
tuplets to be randomized is between two and twenty.
Claim 5
Dependent Claim
21 words
The method of claim 1, wherein the number of
reaction vessels is equal to the number of tuplets to be
randomized.
Claim 6
Dependent Claim
18 words
The method of claim 5, wherein each tuplet
to be randomized is coupled in a separate reaction vessel.
Claim 7
Dependent Claim
21 words
The method of claim 1, wherein the
number of reaction vessels is less than the number of
tuplets to be randomized.
Claim 8
Dependent Claim
22 words
The method of claim 7, wherein two
different tuplets to be randomized are coupled to a support
in a single reaction vessel.
Claim 9
Dependent Claim
29 words
The method of claim 7, wherein the
first, second and third monomers of the tuplet are selected
from (T/G)TT, (T/C)CT, (T/C)AT, (T/C)GT, (C/A)TG, (C/G)AG,
(A/G)CT, (A/G)AT, (T/G)GG or A(T/A)A.
Claim 10
Dependent Claim
49 words
A method of synthesizing
oligonucleotides using individual monomers having at least
one specified tuplet at a predetermined position and the
remaining positions having random tuplets comprising the
method of claim 1, wherein said specified tuplet position
is synthesized on the mixed supports prior to dividing into
separate reaction vessels.
Claim 11
Dependent Claim
21 words
The method of claim 10, wherein the
first position is predetermined and the specified tuplet is
synthesized prior to step (1).
Claim 12
Independent Claim
105 words
A method of synthesizing oligonucleotides
having tuplets which are diverse but biased toward a
predetermined sequence comprising the steps of: (1) sequentially coupling monomers on a
support to form a tuplet with a
predetermined sequence, said coupling
being performed in a first reaction
vessel; (2) sequentially coupling monomers on a
support to form a random tuplet, said
coupling being performed in a second
reaction vessel; (3) mixing the supports from said first and
second reaction vessels; and (4) dividing the mixed supports from step
(3) and using the supports in step (1)
and step (2), wherein steps (1) through
(3) are repeated at least once.
Claim 13
Dependent Claim
15 words
The method of claim 12 further comprising
step (4) cleaving said oligonucleotide from said support.
Claim 14
Dependent Claim
82 words
The method of claim 12, wherein said tuplet
is a triplet and step (2) further comprises: (2a) coupling a mixture of four different
monomers for the first monomer of the
random triplet; (2b) coupling a mixture of four different
monomers for the second monomer of the
random triplet; and (2c) coupling a pair of monomers selected
from the group consisting of guanine
and thymine or cytosine and adenine
monomers for the third monomer of the
random triplet to form a random
triplet.
Claim 15
Dependent Claim
16 words
The method of claim 14, wherein the four
different monomers are adenine, guanine, cytosine and
thymine.
Claim 16
Dependent Claim
19 words
The method of claim 14, wherein four
different monomers in step (2a) or (2b) are present in
equal proportions.
Claim 17
Dependent Claim
20 words
The method of claim 14, wherein the four
different monomers in steps (2a) and (2b) are present in
equal proportions.
Claim 18
Dependent Claim
20 words
The method of claim 14, wherein the four
different monomers in steps (2a) or (2b) are present in
unequal proportions.
Claim 19
Dependent Claim
20 words
The method of claim 14, wherein the four
different monomers in steps (2a) and (2b) are present in
unequal proportions.
Claim 20
Dependent Claim
19 words
The method of claim 14, wherein the guanine
and thymine monomers of step (2c) are present in equal
proportions.
Claim 21
Dependent Claim
19 words
The method of claim 14, wherein the guanine
and thymine monomers of step (2c) are present in unequal
proportions.
Claim 22
Dependent Claim
21 words
The method of claim 12, wherein the mixed
supports from said first and second reaction vessels are
divided into equal proportions.
Claim 23
Dependent Claim
21 words
The method of claim 12, wherein the mixed
supports from said first and second reaction vessels are
divided into unequal proportions.
Claim 24
Dependent Claim
12 words
A diverse population of oligonucleotides
synthesized by the method of claim 1.
Claim 25
Dependent Claim
12 words
A diverse population of oligonucleotides
synthesized by the method of claim 12.
References Cited (Backward Citations)
No citation records indexed.
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