Production of closed linear dna using a palindromic sequence

A primer for the amplification of a DNA template comprising a protelomerase target sequence, particularly for production of closed linear DNA, which primer is capable of specifically binding to a palindromic sequence within a protelomerase target sequence and priming amplification in both directions.

Synthesis apparatus and method

Apparatus for biochemical synthesis comprises a reaction vessel, a temperature control device for the reaction vessel, a plurality of reservoirs for reaction components, and a supply/withdrawal means, e.g. comprising reciprocating syringe pumps and switchable valves. Agitation of a reaction mixture is effected by withdrawing a part of the mixture from the reaction vessel and returning it thereto.

METHOD OF DNA SYNTHESIS

The present invention relates to an in vitro cell-free process for production of deoxyribonucleotides (DNAs) comprising at least one hairpin, corresponding DNA products and uses thereof, and oligonucleotides and kits useful in the process of the invention. 1. An in vitro cell-free process for production of a deoxyribonucleic acid (DNA) molecule which comprises a desired DNA sequence , the process comprising:(a) contacting an oligonucleotide immobilised on a solid support with a series of template oligonucleotides which overlap in sequence, in the presence of at least one DNA polymerase under conditions promoting template-dependent extension of the immobilised oligonucleotide to produce a first DNA strand which comprises the desired DNA sequence and a first portion of a protelomerase target sequence proximal to the solid support;(b) introducing a DNA sequence comprising a second portion of the protelomerase target sequence into a distal end of the first DNA strand, or into a second DNA strand, such that the first and second portions of the protelomerase target sequence thereby create the protelomerase target sequence proximal to the solid support; and(c) contacting the protelomerase target sequence proximal to the solid support with a protelomerase to thereby produce and release the DNA molecule from immobilisation.2. The process according to claim 1 , wherein the immobilised oligonucleotide of (a) comprises the first portion of the protelomerase target sequence.3. The process according to claim 1 , wherein the first portion of the protelomerase target sequence is introduced into the first DNA strand by template-dependent extension.4. The process according to claim 1 , wherein the DNA molecule is a single-stranded circular DNA molecule comprising at least one hairpin that comprises the first portion or the second portion of the protelomerase target sequence.5. The process according to claim 4 , wherein step (b) comprises template-dependent extension with one or more ...

CLOSED LINEAR DNA PRODUCTION

The present invention relates to improved processes for production of closed linear deoxyribonucleic acid (DNA), in particular cell-free enzymatic production of closed linear DNA molecules, preferably using a closed linear DNA as a template for DNA synthesis. The invention further relates to a novel closed linear DNA species, suitable for use as a template in the improved processes for production of closed linear DNA. Further, the invention pertains to the intermediate products of the processes, since this enables the production of larger quantities of closed linear DNA from the template than with methods known in the art. 1. A cell-free method of producing closed linear deoxyribose nucleic acid (DNA) molecules comprising:(a) contacting a template comprising a linear, double stranded DNA molecule covalently closed at each end by a portion of a protelomerase recognition sequence and comprising at least one stem loop motif with a strand-displacing polymerase under conditions promoting amplification of said template in the presence of at least one primer which is capable of binding specifically to a primer binding site within said stem loop motif; and(b) contacting DNA produced in (a) with at least one protelomerase under conditions promoting production of closed linear DNA.2. A method as claimed in wherein a sequence of said stem loop motif comprises a central section flanked by two sequences.3. A method as claimed in wherein the sequence of the central section of the stem loop motif comprises a primer binding site.4. A method as claimed in claim 1 , wherein a sequence forming the stem loop motif is adjacent or near to the portion of the protelomerase recognition sequence within the covalently closed end of the template.5. A method as claimed in claim 1 , wherein a sequence of said stem loop motif is capable of forming a stem loop structure under conditions promoting the formation of a secondary structure.6. A method as claimed in wherein the sequence of the stem loop ...

Macrolide antibiotics and their preparation

NOVEL ANTIBIOTIC COMPOUND AND METHODS FOR PREPARING THE SAME

L'INVENTION CONCERNE UN NOUVEAU COMPOSE REPONDANT A LA FORMULE I CI-DESSOUS. CE COMPOSE POSSEDE UNE ACTIVITE ANTIPARASITE ET CONSTITUE UN INTERMEDIAIRE INTERESSANT POUR LA PREPARATION D'AUTRES COMPOSES A ACTIVITE ANTIBIOTIQUE. L'INVENTION CONCERNE EN OUTRE DES PROCEDES POUR LA PREPARATION DU COMPOSE DE LA FORMULE I.

Antibiotic compounds, their preparation and the pharmaceutical compositions which contain them

L'INVENTION CONCERNE DE NOUVEAUX MACROLIDES. CES COMPOSES REPONDENT A LA FORMULE I (CF DESSIN DANS BOPI) ET LEURS SELS, OU R METHYLE, ETHYLE OU ISOPROPYLE; R REPRESENTE H, ALKYLE EN C-C OU ALCENYLE EN C-C ET NOR EST EN CONFIGURATION E; OR EST HYDROXYLE OU HYDROXYLE SUBSTITUE AVEC JUSQU'A 25ATOMES DE CARBONE. CES COMPOSES SONT INTERESSANTS POUR LUTTER CONTRE DES INSECTES, DES ACARIENS, DES NEMATODES OU D'AUTRES PARASITES.

A process for the preparation of therapeutically useful macrolide compounds

Biochemical synthesis apparatus

A biochemical synthesis apparatus comprises a receptacle for containing a medium, and a support on which a microorganism can be placed. The support can be placed in contact with the medium, so as to allow access of the microorganism to the medium. The support with the microorganism can be removed from the medium, allowing the medium to be replaced. By this, the receptacle can firstly contain a growth medium, and secondly a secondary medium capable of causing the production by the microorganism of a potentially useful biochemical.

Macrolide derivatives.

Process for the preparation of macrolide compounds

A process for the preparation of a compound of formula (I)

Macrolide compounds

Abstract: Compounds are described of the formula (I) and salts thereof, wherein R1 is methyl, ethyl or isopropyl; R2 is -H, or a group -OR5 (wherein -OR5 is a hydroxyl group or a substituted hydroxyl group having up to 25 carbon atoms); and R3 is -H, or R2 and R3 together with the carbon atom to which they are attached represent C=CH2, C=O or C=NOR6 (where R6 is -H, alkyl or alkenyl and the C=NOR6 is in the E configuration); OR4 is as defined above for OR5 and one of the symbols X represents an epoxide oxygen atom and the other represents an epoxide oxygen atom or a carbon-carbon bond. These compounds may be used for controlling insect, acarine, nematode or other pests.

MACROLIDE COMPOUNDS, PROCEDURES FOR PREPARING THEREOF, PREPARATIONS CONTAINING SUCH COMPOUNDS, AND NON-THERAPEUTIC PROCEDURE TO COMPLETE PESTS USING USE

ANTIBIOTIC MACROLIDE COMPOUNDS, PREPARATIONS CONTAINING THESE, PROCEDURES FOR THE PREPARATION OF THE COMPOUNDS, AND PROCEDURES FOR USING THE COMPOUNDS OR PREPARATIONS

NOVEL ANTIBIOTIC COMPOUNDS AND PREPARATION METHOD

Macrolide antibiotics

Compounds are described of formula (I) ##STR1## and salts thereof, wherein OR is a hydroxyl group or a substituted hydroxyl group having up to 25 carbon atoms, R 1 represents a methyl, ethyl or isopropyl group and the broken line represents a double bond at the 22,23-position or at the 23,24-position. These compounds may be used for controlling insect, acarine, nematode or other pests.

Process for preparing macrolide compounds with antibiotic activity

Macrolide compounds

PROCEDURE FOR OBTAINING AN ANTIBIOTIC

Macrolide compounds

A B S T R A C T MACROLIDE ANTIBIOTICS Compounds are described of formula (I) (I) and salts thereof, wherein R1 represents a methyl, ethyl or isopropyl group; R2 is alkyl; alkyl substituted by carboxy or alkoxycarbonyl; alkenyl; phenyl or phenalkyl; R3 is -OH; OR4 is a hydroxyl group or a substituted hydroxyl group having up to 25 carbon atoms. These compounds may be used for controlling insect, acarine, nematode or other pests.

Macrolide compounds

A B S T R A C T Compounds are described of formula (I) (I) and salts thereof, wherein R1 a methyl, ethyl or isopropyl group; R2 is -H, -OH, substituted -OH or a phosphate ester group) and R3 is -H, or R2 and R3 together with the carbon atom to which they are attached represent a >C=0 group and OR4 is a group OR5 as defined above; with the proviso that either R2 represents a group OR5 (where OR5 is an alkoxy group OR7 in which R7 represents a C1-6 alkyl substituted by a halogen atom or C2-6 alkyl interrupted by -0- or -S-) or at least one of R2 and OR4 is a phosphate ester group or a group OCONR10R11 (where R10 and R11 are -H or C1-4 alkyl). These compounds may be used for controlling insect, acarine, nematode or other pests.

Macrolide compounds

ABSTRACT Compounds of formula (I) (I) and salts thereof wherein R1 is a methyl, ethyl or isopropyl group; R2 is a hydrogen atom or a group OR4 (where OR4 is a hydroxyl group or a substituted hydroxyl group having up to 25 carbon atoms) and R3 is a hydrogen atom, or (when R2 is a hydrogen atom) a bromine atom or R2 and R3 together with the carbon atom to which they are attached represent >C=CH2, >C=0 or >C=NOR5 (where R5 is a hydrogen atom, a C1-8 alkyl group or a C3-8 alkenyl group and the group >C-NOR5 is in the E configuration); and Y represents an oxygen atom or a group NOR5 (where R5 is as defined above); with the proviso that when Y is an oxygen atom and R1 is an isopropyl group, then R2 is other than hydroxyl group. These compounds may be used for controlling insect, acarine, nematode or other pests.

Macrolide compounds

Macrolide compounds

Compounds of formula (I) <CHEM> and salts thereof, wherein R is a C1-4 alkyl group, R<1> is a methyl, ethyl or isopropyl group; and OR<2> is a hydroxyl or substituted hydroxyl group having up to 25 carbon atoms. These compounds may be used for controlling insect, acarine, nematode or other pests.

Macrolide compounds

Compounds of formula (I): and salts thereof; wherein R 1 is -H, -OH or substituted -OH and R 2 is -H, or R 1 and R 2 together with the carbon atom to which they are attached represent C=0, C=CH 2 or C=NOR 6 in the E configuration (where R 6 is -H, alkyl or alkenyl); R 3 is -OH or substituted -OH and R 4 is -H, or R 3 and R 4 together with the carbon atom to which they are attached represent C=0; -A-B- is -C(CH 3 ) = CH- or -C(=CH 2 )CHX- (where Y is Cl or Br) and Y is -C(R 8 )=CHR 7 - or -C(=CH 2 )CHXR 7 (where X is Cl or Br, R 7 is methyl, ethyl or isopropyl and R 8 is C 2 - 8 alkyl or C 7 - 15 phenalkyl); with the proviso that when -A-B-is(=CH 2 )CHX-, then Y is -C(= CH2 )C HXR 7. These compounds may be used for controlling insect, acarine, nematode or other pests.

Synthesis apparatus and method

PROCESS FOR THE PREPARATION OF A COMPOUND

Process for preparation of macrolides

Pesticide

PREPARATION OF CLOSED, LINEAR DNA

Closed linear dna production

Synthesis of dna with improved yield

The present invention relates to an improved process for synthesis of deoxyribonucleic acid (DNA), in particular cell-free enzymatic synthesis of DNA, preferably on a large scale, with an improved yield and/or with an improved efficiency. The species of cation present in the nucleotide salt as the counter-ion is critical to the yield, efficiency and fidelity of high yielding enzymatic DNA synthesis reaction. The processes herein use alternative cations as counter-ions for the ionic nucleotides, permitting the use of higher concentrations of nucleotides in DNA synthesis, and further allowing for more favourable reaction conditions to be used.

Macrolide compounds

Compounds are described oftheformula and salts thereof, wherein R 1 is methyl, ethyl or isopropyl; R 2 is-H,-OH orsubstituted-OH and R 3 is-H, or R 2 and R 3 together with the carbon atom to which they are attached represent C=CH 2 , C=O or C=NOR 6 (where R 6 is-H, alkyl or alkenyl and the C=NOR 6 is inthe E configuration); OR 4 is as defined above for OR 5 and one of the symbols X represents an epoxide oxygen atom and the other represents an epoxide oxygen atom or a carbon-carbon bond. These compounds maybe used for controlling insect, acarine, nematode or other pests.

Macrolide compounds

ABSTRACT MACROLIDE COMPOUNDS Compounds are described of formula (1) (1) and salts thereof wherein R1 is a methyl, ethyl or isopropyl group; R2 represents a hydrogen atom ox a group OR4 (where OR4 is a hydroxyl group or a substituted hydroxyl group having up to 25 carbon atoms) and R3 represents a hydrogen atom, or R2 and R3 together with the carbon atom to which they are attached represent >C=CH2, >C=0 or >C=NOR5 (where R5 is a hydrogen atom or a C1-8 alkyl group and the group >C=NOR5 is in the E configuration). These compounds may be used for controlling insect, acarine, nematode or other pests.

Pesticide means and method for control of pests

The agent and the method are applied in agriculture and veterinary medicine. The agent is less toxic with regard to the warm-blooded animals. Along with the usual additives, it contains a pesticide-effective quantity from any of the compounds with a common formula:<IMAGE>,where R1 means methyl, ethyl- or isopropyl residue, while R2 is hydrogen or methyl radical.

Process for production of macrolide compounds and pharmaceutical preparations containing them as well as pesticidal preparations

Macrolide compounds

Compounds of formula (I) <CHEM> and salts thereof, wherein R is a C1-4 alkyl group, R<1> is a methyl, ethyl or isopropyl group; and OR<2> is a hydroxyl or substituted hydroxyl group having up to 25 carbon atoms. These compounds may be used for controlling insect, acarine, nematode or other pests.

Macrolide antibiotics and their preparation

ABSTRACT Compounds are described of formula (I)

Macrolide compounds

MACROLID COMPOUNDS.

Compounds are described of formula (I) <CHEM> and salts thereof, wherein R<1> represents a methyl, ethyl or isopropyl group; R<2> is alkyl; alkyl substituted by carboxy or alkoxycarbonyl; alkenyl; phenyl or phenalkyl; R<3> is-OH; OR<4> is a hydroxyl group or a substituted hydroxyl group having up to 25 carbon atoms. These compounds may be used for controlling insect, acarine, nematode or other pests.

Synthesis of dna with improved yield.

La presente invención se refiere a un proceso mejorado para la síntesis de ácido desoxirribonucleico (ADN), en particular una síntesis enzimática de ADN libre de células, preferiblemente a gran escala, con un rendimiento mejorado y/o con una eficiencia mejorada. La especie de catión presente en la sal de nucleótido como el contraión es crucial para el rendimiento, eficiencia y fidelidad de la reacción de síntesis enzimática de ADN de alto rendimiento. Los procesos descritos en este documento usan cationes alternativos como contraiones para los nucleótidos iónicos, lo que permite el uso de concentraciones más altas de nucleótidos en la síntesis de ADN, y permite además que se usen condiciones de reacción más favorables.

Macrolide antibiotics and their preparation

Compounds are described of formula (1)and salts thereof, wherein OR is a hydroxyl group or a substituted hydroxyl group having up to 25 carbon atoms, R¹ represents a methyl, ethyl or isopropyl group and the broken line represents a double bond at the 22,23-position or at the 23,24-position.These compounds may be used for controlling insect, acarine, nematode or other pests.

ANTIBIOTIC COMPOUNDS, ANTI-PARASITARY COMPOSITIONS THAT CONTAIN THEM AND PROCEDURE TO PREPARE AND APPLY THEM

Macrolide antibiotics and their preparation

A B S T R A C T MACROLIDE ANTIBIOTICS AND THEIR PREPARATION Compounds are described of formula (I) (I) and salts thereof, wherein R1 represents a methyl, ethyl or isopropyl group; and OR2 is a hydroxyl group or a substituted hydroxyl group having up to 25 carbon atoms. These compounds may be used for controlling insect, acarine, nematode or other pests.

Macrolid compounds

Compounds of formula (I): ##STR1## and salts thereof; wherein R 1 is --H, --OH or substituted --OH and R 2 is --H, or R 1 and R 2 together with the carbon atom to which they are attached represent C═O, C═CH 2 or C═NOR 6 in the E configuration (where R 6 is --H, alkyl or alkenyl); R 3 is --OH or substituted --OH and R 4 is --H, or R 3 and R 4 together with the carbon atom to which they are attached represent C═O; --A--B-- is --C(CH 3 )═CH-- or --C(═CH 2 )CHX-- (where X is Cl or Br) and Y is --C(R 8 )═CHR 7 -- or --C(═CH 2 )CHXR 7 (where X is Cl or Br, R 7 is methyl, ethyl or isopropyl and R 8 is C 2-8 alkyl or C 7-15 phenalkyl); with the proviso that when --A--B-- is (═CH 2 )CHX--, then Y is--C(C═CH 2 )CHXR 7 . These compounds may be used for controlling insect, acarine, nematode or other pests.

Chemical compounds and their use as pesticides

Macrolide antibiotics and their preparation

Macrolide compounds

ANTIBIOTIC COMPOUNDS, THEIR PREPARATION AND THE PHARMACEUTICAL COMPOSITIONS CONTAINING THEM

Biochemical synthesis apparatus

The apparatus comprises a receptacle 10, for nutrient medium 18, having a lid 12 with a gas-permeable foam insert 16 and a floating raft 20 carrying a fermentation vessel 28 for a microorganism which is in contact with the growth medium via a membrane (32) of the vessel. The supporting raft and vessel can be removed so that the medium can be replaced by a second medium which provides conditions for biosynthesis of a potentially useful biochemical, such as a secondary metabolite, by the microorganism. In a further embodiment, the apparatus comprises a cylindrical receptacle and a cylindrical fermentation vessel having a tapered end which has membranes formed thereacross (Figs.12,13). Alternatively, the vessel is formed with an outer membrane extending down its length (Figs.15,16).

A PROCEDURE FOR PREPARING MACROLID ANTIBIOTICS AND THEIR USE.

SE DESCRIBEN COMPUESTOS DE FORMULA (I) Y SALES DE LOS MISMOS, EN LA QUE R1 REPRESENTA UN GRUPO METILO, ETILO O ISOPROPILO; Y OR2 ES UN GRUPO HIDROXILO O UN GRUPO HIDROXILO SUSTITUIDO QUE TENGA HASTA 25 ATOMOS DE CARBONO. ESTOS COMPUESTOS PUEDEN USARSE PARA CONTROLAR PLAGAS DE INSECTOS, ACAROS, NEMATODOS Y OTROS. COMPOUNDS OF FORMULA (I) AND SALTS OF THEM ARE DESCRIBED, IN WHICH R1 REPRESENTS A METHYL, ETHYL OR ISOPROPYL GROUP; AND OR2 IS A HYDROXYL GROUP OR A SUBSTITUTED HYDROXYL GROUP THAT HAS UP TO 25 CARBON ATOMS. THESE COMPOUNDS MAY BE USED TO CONTROL PESTS OF INSECTS, MITES, NEMATODES AND OTHERS.

Macrolide compounds

Compounds are described of formula (I) ##STR1## and salts thereof, wherein R 1 represents a methyl, ethyl or isopropyl group; R 2 represents a hydrogen atom, a C 1-8 alkyl group or a C 3-8 alkenyl group and the group ═NOR 2 is in the E-configuration; OR 3 is a hydroxyl group or a substituted hydroxyl group having up to 25 carbon atoms. These compounds may be used for controlling insect, acarine, nematode or other pests.

Process for preparing macrolide compounds and pharmaceuticals comprising the same

ANTIBIOTICS

Macrolide compounds

A B S T R A C T MACROLIDE ANTIBIOTICS Compounds are described of the formula (I) (I) and salts thereof, wherein R1 is a methyl, ethyl or isopropyl group; R2 is -H, -OH or substituted -OH and R3 is -H, or R2 and R3 together with the carbon atom to which they are attached represent >C=0, >C=CH2 or >C=NOR7 (where R7 is -H, alkyl, alkenyl and the group >C=NOR is in the E configuration); OR4 is -OH or substituted -OH and R6 is -H or a C1-4 alkanoyl group. These compounds may be used for controlling insect, acarine, nematode or other pests.

Macrolide compounds

COMPOUNDS ACTIVE AGAINST PEST, COMPOSITION FOR PEST CONTROL, PROCESS TO COMBAT PEST, AND PROCESS TO PREPARE THE COMPOUND

ANTIBIOTIC MACROLIDE COMPOUNDS, PREPARATIONS CONTAINING THESE, PROCEDURES FOR THE PREPARATION OF THE COMPOUNDS, AND PROCEDURES FOR USING THE COMPOUNDS OR PREPARATIONS

Milbemycin derivatives and compositions

Antiparassitarie che li contengono composti antibiotici, composizioni e procedimento per prepararli ed applicarli

Menetelmä terapeuttisesti käyttökelpoisten makrolidiyhdisteiden valmis tamiseksi

Composti antibiotici macrolidi, composizioni che li contengono e procedimento per prepararli ed applicarli come pesticidi

Composto antibiotico, composizioni che lo contengono e relativo metodo di preparazione e applicazione

DNA:n synteesi

Synthesis of dna

Production of closed linear dna using a palindromic sequence

A primer for the amplification of a DNA template comprising a protelomerase target sequence, particularly for production of closed linear DNA, which primer is capable of specifically binding to a palindromic sequence within a protelomerase target sequence and priming amplification in both directions.

Dnr sintezė

Dna-syntese

Antibiotic compounds

Apparatus for preserving microorganisms

Subculturing apparatus (30) comprises a receptacle for supporting a growth medium (5) for culturing of a microorganism. The microorganism is entrained to grow in a predetermined direction, towards an end of the growth medium (50), where a further apparatus (30) can be placed adjacent thereto. The microorganism can then grow into the growth medium of the further apparatus. The method of storing a microorganism for use in microbiological processes is also described, as is a method of fermentation of a stored microorganism for the production of biochemicals such as pharmaceuticals or agrochemicals.

Dna synthesis yield improvements

The present invention relates to an improved process for synthesis of deoxyribonucleic acid (DNA), in particular cell-free enzymatic synthesis of DNA, preferably on a large or industrial scale, with an improved yield and/or with an improved efficiency. The invention requires the use of nucleotide complexes wherein the nucleotide is associated with a mixture of divalent and monovalent cations. Preferably, the divalent cation may be magnesium or manganese.

Dna synthesis yield improvements

The present invention relates to an improved process for synthesis of deoxyribonucleic acid (DNA), in particular cell-free enzymatic synthesis of DNA, preferably on a large or industrial scale, with an improved yield and/or with an improved efficiency. The invention requires the use of nucleotide complexes wherein the nucleotide is associated with a mixture of divalent and monovalent cations. Preferably, the divalent cation may be magnesium or manganese.

Dna synthesis yield improvements

The present invention relates to an improved process for synthesis of deoxyribonucleic acid (DNA), in particular cell-free enzymatic synthesis of DNA, preferably on a large or industrial scale, with an improved yield and/or with an improved efficiency. The invention requires the use of nucleotide complexes wherein the nucleotide is associated with a mixture of divalent and monovalent cations. Preferably, the divalent cation may be magnesium or manganese.

Procédé d'oxydation d'hydrazoformamides

Makrolid-verbindungen.

Production of closed linear dna using a palindromic sequence

Makrolide antibiotika und verfahren zu deren herstellung.

Verfahren zur Herstellung von Azodicarbonamiden

Synthesis of dna

The present invention relates to an improved process for synthesis of DNA, RNA, proteins and like molecules, in particular cell-free enzymatic synthesis of DNA, preferably in large scale. The present invention relates to the synthesis of DNA using strand-displacement replication and the addition of nucleotides to the reaction mixture is controlled, thus controlling yield. The reaction mixture contains a starting amount of nucleotides, polymerase and DNA template, to which further nucleotides are supplied in a controlled manner.

Síntesis del ADN

La presente invención se refiere a un proceso mejorado para la síntesis de ADN, ARN, proteínas y moléculas similares, en particular la síntesis enzimática de ADN sin células, preferiblemente a gran escala. La presente invención se refiere a la síntesis de ADN mediante replicación por desplazamiento de cadena y se controla la adición de nucleótidos a la mezcla de reacción, controlando así el rendimiento. La mezcla de reacción contiene una cantidad inicial de nucleótidos, polimerasa y molde de ADN, a los que se suministran más nucleótidos de forma controlada. (Traducción automática con Google Translate, sin valor legal)

Production of closed linear DNA using a palindromic sequence

A primer for the amplification of a DNA template comprising a protelomerase target sequence, particularly for production of closed linear DNA, which primer is capable of specifically binding to a palindromic sequence within a protelomerase target sequence and priming amplification in both directions.