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CiteWeb id: 19830000003. CiteWeb score: 27848. DOI: 10.1016/0003-2697(83)90418-9. A technique for conveniently radiolabeling DNA restriction endonuclease fragments to high specific activity is described. DNA fragments are purified from agarose gels directly by ethanol precipitation and are then denatured and labeled with the large fragment of DNA polymerase I, using random oligonucleotides as primers. Over 70% of the precursor triphosphate is routinely incorporated into complementary DNA, and specific activities of over 109 dpm/μg of DNA can be obtained using relatively small amounts of precursor. These "oligolabeled" DNA fragments serve as efficient probes in filter hybridization experiments.. Links: ...
TY - JOUR. T1 - Inhibition of sequence-specific protein-DNA interaction and restriction endonuclease cleavage via triplex stabilization by poly(L-lysine)-graft-dextran copolymer. AU - Ferdous, Anwarul. AU - Akaike, Toshihiro. AU - Maruyama, Atsushi. PY - 2000/6. Y1 - 2000/6. N2 - Triplex stabilization by poly(L-lysine)-graft-dextran copolymer within a mammalian gene promoter inhibits the DNA binding activity of nuclear proteins from HeLa cells as well as restriction endonuclease cleavage at physiological pH and ionic conditions in vitro. Electrophoretic mobility shift assays using a 30-mer hornopurine-homopyrimidine stretch (located between -170 and -141 bp) of rat α1 (I) collagen gene promoter reveal that the copolymer, at its wide range of charge ratio with DNA, stabilizes triplex DNA and enhances triplex-specific inhibition of the protein - DNA interaction. When the triplex-forming region (located between -165 and -146 bp) of the promoter is engineered at the Bam H1 and Pst 1 sites of a ...
Define restriction endonuclease. restriction endonuclease synonyms, restriction endonuclease pronunciation, restriction endonuclease translation, English dictionary definition of restriction endonuclease. Noun 1. restriction endonuclease - any of the enzymes that cut nucleic acid at specific restriction sites and produce restriction fragments; obtained from...
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DNA Restriction Enzymes from Takara such as BmeT110I are high-quality: perform restriction enzyme digestion with reliable restriction endonucleases
DNA Restriction Enzymes from Takara such as HpaI are high-quality: perform restriction enzyme digestion with reliable restriction endonucleases
Restriction enzymes are also called molecular scissors as they cleave DNA at or near specific recognition sequences known as restriction sites. These enzymes make one incision on each of the two strands of DNA and are also called restriction endonucleases.4, 5. Viruses infect the host cells by injecting their DNA into the cells. This viral DNA hijacks the host cells machinery for reproduction of viral progeny, resulting in the host cells death. To overcome the viral infection, many bacteria and archaea have evolved several mechanisms. A major protective mechanism involves the use of restriction enzymes to degrade the invading viral DNA by cleaving it at specific restriction sites. At the same time, the host cell protects its own DNA from being cleaved by employing other enzymes called methylases, which methylate adenine or cytosine bases within host recognition sequences. For each of the restriction enzyme, the host cell produces a corresponding methylase that methylates and protects the ...
Restriction Enzyme Digest not working - posted in Molecular Cloning: Hello,For some reason, my digestions are not working. I am afraid to consult my PI because the cause of the problem might be something that has to do with my procedures rather the actual supplies that I am using. I hope someone can pinpoint what I am doing wrong. Thank you!Experiment goal:1. Create insert with sticky ends by introducing restriction sites (Kpnl and Xbal) through PCR, then digesting those ends (with Kpn...
A BioBrick is a sequence of DNA with a predefined structure and function. This "payload" is held in a circular plasmid, which is an isolated, circular piece of DNA that can replicate in bacteria. BioBricks™ are created with the intention of being easily joined and manipulated. In order for this to be possible, the BioBrick™ assembly standard requires the use of defined prefix and suffix sequences (flanking both sides of the BioBrick) that contain specific restriction endonuclease sites. These sites are called EcoRI, NotI and XbaI in the upstream, and SpeI, NotI, and PstI in the downstream. Naturally, the parts must also be engineered such that these sites are not present in the functional region of the sequence.[3] Cutting the BioBrick at specific restriction sites (using restriction enzymes) is what gives a BioBrick its interlocking ends. The end of one BioBrick can then be connected, or ligated, together with the end of another BioBrick, allowing you to effectively string together ...
A BioBrick is a sequence of DNA with a predefined structure and function. This "payload" is held in a circular plasmid, which is an isolated, circular piece of DNA that can replicate in bacteria. BioBricks™ are created with the intention of being easily joined and manipulated. In order for this to be possible, the BioBrick™ assembly standard requires the use of defined prefix and suffix sequences (flanking both sides of the BioBrick) that contain specific restriction endonuclease sites. These sites are called EcoRI, NotI and XbaI in the upstream, and SpeI, NotI, and PstI in the downstream. Naturally, the parts must also be engineered such that these sites are not present in the functional region of the sequence.[3] Cutting the BioBrick at specific restriction sites (using restriction enzymes) is what gives a BioBrick its interlocking ends. The end of one BioBrick can then be connected, or ligated, together with the end of another BioBrick, allowing you to effectively string together ...
FlyCut® BamHI,Fast Restriction Enzyme,Restriction Enzyme and Modification Enzyme,Products,Beijing TransGen Biotech Co.Ltd,OverviewContents& storageCitations & referencesRelated ImagesDownloadOverviewDescriptionFlyCut ® BamHI is expressed and
This was a hybrid restriction site created during the cloning process (Cap-trapper) for the library creation. Neither of these restriction sites will work to digest the insert from the vector. According to the IMAGE consortium the SalI-XhoI (gtcgag) is located at position 742 of the polylinker sequence (http: //mgc. nci. nih. gov/Vectors/vec_pbluescriptr). Customers can use BamHI (5) and EcoRI (3) to digest out the insert. Other options for sub-cloning are either to use different restriction enzymes lying outside BamHI and EcoRI or to design insert-specific primers based on the insert sequence. You can also find a reference for the cap trapper method here: Carninci et. al. , DNA RESEARCH 4, 61-66 (1997), High Efficiency Selection of Full-length cDNA by Improved Biotinylated Cap Trapper ...
無法找到符合 plasmid dna preparation restriction enzyme digestion and electrophoresis 的相關結果。請嘗試以下建議或輸入其它關鍵字。 ...
Restriction enzymes BamH1 and pst1, in pMA has the same weight(1.2kb) with one of the fragment restricted by ECORI and pst1 in pMB.. The size of the restriction fragments of the plasmids pMA and pMB when digested with the enzymes was roughly obtained with the help of 1kb marker and these values were not accurate. As the pst1 has one restriction site in the pMA and two restriction sites in the pMB, we can assume that pMA is part of pMB. If the longer fragment of the pst1 site in the pMB when re-circled will have the same restriction enzymes sites as the pMA(BamH1,ECORI,pst1). The xhol restriction site in the pMB is not in the longer fragment and so this xhol site will not digest the longer fragment which is consistent with pMA.. The strains of DH5?, PUC19, pMA, pMB, and XL1-blue are tested for antibiotic resistance in Luria-Broth agar plates and the results noted. The bacterial host, DH5? has no resistance to any of the three bacterial strains, pUC19 is resistant to ampicillin, pMA is resistant ...
A restriction endonuclease having one DNA binding site is proposed, synthesized from a restriction endonuclease that has one C-terminal domain and one N-terminal domain and two DNA binding sites, by proteolytic cleavage into the two domains or by cloning the gene segment that codes for the domains and expression of the domains and selection of the endonucleolytic domains having one DNA binding site. In addition, a method of synthesis of the restriction endonuclease and its use are claimed.
Edvotek. Analysis of Eco RI Cleavage Patterns of Lambda DNAThis experiment introduces the use of restriction enzymes as a tool to digest DNA at specific nucleotide sequences. …
References[edit] ^ Roberts RJ (Nov 1976). "Restriction endonucleases". CRC Critical Reviews in Biochemistry. 4 (2): 123-64. doi:10.3109/10409237609105456. PMID 795607. ^ a b Kessler C, Manta V (Aug 1990). "Specificity of restriction endonucleases and DNA modification methyltransferases a review (Edition 3)". Gene. 92 (1-2): 1-248. doi:10.1016/0378-1119(90)90486-B. PMID 2172084. ^ Pingoud A, Alves J, Geiger R (1993). "Chapter 8: Restriction Enzymes". In Burrell M. Enzymes of Molecular Biology. Methods of Molecular Biology. 16. Totowa, NJ: Humana Press. pp. 107-200. ISBN 0-89603-234-5. ^ a b Arber W, Linn S (1969). "DNA modification and restriction". Annual Review of Biochemistry. 38: 467-500. doi:10.1146/annurev.bi.38.070169.002343. PMID 4897066. ^ Krüger DH, Bickle TA (Sep 1983). "Bacteriophage survival: multiple mechanisms for avoiding the deoxyribonucleic acid restriction systems of their hosts". Microbiological Reviews. 47 (3): 345-60. PMC 281580 . PMID 6314109. ^ Kobayashi I (Sep 2001). ...
Based on plasmid maps provided, students will choose restriction enzymes and perform a restriction enzyme digest that will allow them to identify two unknown plasmids. ...
NEB scientists continue to improve our portfolio of restriction enzymes, as well as explore their utility in new technologies.
Molecular Cloning, also known as Maniatis, has served as the foundation of technical expertise in labs worldwide for 30 years. No other manual has been so popular, or so influential.
In the following sequence find restriction sites for EcoRI, HindIII and Hae III. Show how many fragments will be produced by restriction by all of these enzymes at the same time and their size, Which of the fragments produced.
Linear) (Six-base) MAPSORT of: KIEE0945.seq Check: 8372 from: 1 to: 1050 KIEE0945.orf With 182 enzymes: SgfI * April 24, 2009 13:19 .. AcuI CTGAAGnnnnnnnnnnnnnn_nn Cuts at: 0 162 271 645 1050 Size: 162 109 374 405 Fragments arranged by size: 405 374 162 109 AflIII ACryG_T Cuts at: 0 396 518 1050 Size: 396 122 532 Fragments arranged by size: 532 396 122 AlfI GCAnnnnnnTGCnnnnnnnnnn_nn Cuts at: 0 329 1050 Size: 329 721 AloI GAACnnnnnnTCCnnnnnnn_nnnnn Cuts at: 0 694 726 1050 Size: 694 32 324 Fragments arranged by size: 694 324 32 AlwNI CAG_nnnCTG Cuts at: 0 375 645 1050 Size: 375 270 405 Fragments arranged by size: 405 375 270 AseI ATTA_AT Cuts at: 0 566 1050 Size: 566 484 AvaI CyCGr_G Cuts at: 0 41 1050 Size: 41 1009 BanI GGyrC_C Cuts at: 0 48 1050 Size: 48 1002 BbsI GAAGACnnnnnn_ Cuts at: 0 634 1050 Size: 634 416 BciVI GTATCCnnnnn_n Cuts at: 0 999 1050 Size: 999 51 BclI TGATC_A Cuts at: 0 74 1050 Size: 74 976 BglI GCCn_nnnnGGC Cuts at: 0 590 1050 Size: 590 460 Bme1580I G_kGCmC Cuts ...
What I am confused by though, is the 1/2 and 1/4 component of the answer given. Can anyone give me a breakdown of that please ...
Is this valid, I basically have a type which contains a sequence of itself. Then I define two restrictions, Blue and Green, and then comes my question.... called MyQuestionBase, can I substitute the unbounded item element with just two subclasses Blue and Green. This seems like substitution groups, but they are not global elements, they are global complex types that I am substituting for item in the restriction. This validates OK, but I dont really trust if it is valid????? Please help joey coyle ,?xml version=1.0 encoding=UTF-8?, ,xs:schema xmlns:xs=http://www.w3.org/2001/XMLSchema elementFormDefault=qualified attributeFormDefault=unqualified, ,xs:complexType name=Base, ,xs:sequence, ,xs:element name=item type=Base maxOccurs=unbounded/, ,/xs:sequence, ,/xs:complexType, ,xs:complexType name=BlueBase, ,xs:complexContent, ,xs:restriction base=Base, ,xs:sequence, ,xs:element name=item type=Base maxOccurs=unbounded/, ,/xs:sequence, ,/xs:restriction, ...
Rescue of the ABI3 coding region into a Gateway Entry vector. (A) Schematic diagram showing the location of primers and restriction enzyme sites used for analys
For turn restrictions whose via member is a way: full in-editor support does not exist. You will need to create and edit relations of this type by manually creating the Restriction relation. Highlight the way which will be the via member, create a new Restriction relation with the restriction type you intend, and label the way as a via member in this new relation. Lastly, add the ways which will be the from and to way members, labeling each accordingly with their relation membership. Be careful not to confuse the turn restriction you are working on with others in the same location! Use a temporary name for the new relation (you can delete this name when finished adding members) if this helps you keep track ...
Linear) (Six-base) MAPSORT of: KIAA0030.seq Check: 2035 from: 1 to: 2712 KIAA0030.orf With 182 enzymes: SgfI * March 3, 2008 10:54 .. AarI CACCTGCnnnnnnnn_ Cuts at: 0 869 2517 2712 Size: 869 1648 195 Fragments arranged by size: 1648 869 195 AatII G_ACGTC Cuts at: 0 2372 2712 Size: 2372 340 AccI GTmk_AC Cuts at: 0 620 2712 Size: 620 2092 AcuI CTGAAGnnnnnnnnnnnnnn_nn Cuts at: 0 2179 2527 2640 2712 Size: 2179 348 113 72 Fragments arranged by size: 2179 348 113 72 AfeI AGCGCT Cuts at: 0 2100 2712 Size: 2100 612 AflIII ACryG_T Cuts at: 0 662 2123 2712 Size: 662 1461 589 Fragments arranged by size: 1461 662 589 AhdI GACnn_nnnGTC Cuts at: 0 320 2712 Size: 320 2392 AlfI GCAnnnnnnTGCnnnnnnnnnn_nn Cuts at: 0 2359 2712 Size: 2359 353 ApaI G_GGCCC Cuts at: 0 185 571 1725 2559 2712 Size: 185 386 1154 834 153 Fragments arranged by size: 1154 834 386 185 153 ApoI rAATT_y Cuts at: 0 1762 2712 Size: 1762 950 AvrII CCTAG_G Cuts at: 0 168 2712 Size: 168 2544 BaeI ACnnnnGTAyCnnnnnnn_nnnnn Cuts at: 0 ...
High Fidelity (HF) Restriction Enzymes have 100% activity in CutSmart Buffer; single-buffer simplicity means more straightforward and streamlined sample processing. HF enzymes also exhibit dramatically reduced star activity
Question - It seems that there has developed some restriction to my inserting - NH. Find the answer to this and other Medical questions on JustAnswer
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Isolated restriction enzymes are used to manipulate DNA for different scientific applications. They are used to assist insertion of genes into plasmid vectors during gene cloning and protein production experiments. For optimal use, plasmids that are commonly used for gene cloning are modified to include a short polylinker sequence (called the multiple cloning site, or MCS) rich in restriction enzyme recognition sequences. This allows flexibility when inserting gene fragments into the plasmid vector; restriction sites contained naturally within genes influence the choice of endonuclease for digesting the DNA, since it is necessary to avoid restriction of wanted DNA while intentionally cutting the ends of the DNA. To clone a gene fragment into a vector, both plasmid DNA and gene insert are typically cut with the same restriction enzymes, and then glued together with the assistance of an enzyme known as a DNA ligase.[60][61] Restriction enzymes can also be used to distinguish gene alleles by ...
A restriction map is a map of known restriction sites within a sequence of DNA. Restriction mapping requires the use of restriction enzymes. In molecular biology, restriction maps are used as a reference to engineer plasmids or other relatively short pieces of DNA, and sometimes for longer genomic DNA. There are other ways of mapping features on DNA for longer length DNA molecules, such as mapping by transduction. One approach in constructing a restriction map of a DNA molecule is to sequence the whole molecule and to run the sequence through a computer program that will find the recognition sites that are present for every restriction enzyme known. Before sequencing was automated, it would have been prohibitively expensive to sequence an entire DNA strand. To find the relative positions of restriction sites on a plasmid, a technique involving single and double restriction digests is used. Based on the sizes of the resultant DNA fragments the positions of the sites can be inferred. Restriction ...
This is an interface to a utility that reports all intervals of a desired size between cutting sites of either a single restriction enzyme, or a pair of enzymes. All cutting sites are determined in the human sequence. Error tolerance for interval sizes is selectable, as well as the direction of matching for the determination of enzyme cutting sites. First enzyme (mandatory): AatII AccI Acc65I AciI AflII AflIII AgeI AluI AlwI AlwNI ApaI ApaLI ApoI AscI AseI AvaI AvaII AvrII BamHI BanI BanII BbsI BbvI BcgI BcgI(second_site) BclI BfaI BglI BglII BpmI Bpu1102I BsaI BsaAI BsaBI BsaHI BsaJI BsaWI BsgI BsiEI BsiHKAI BsiWI BslI BsmI BsmAI BsmFI Bsp120I Bsp1286I BspDI BspEI BspHI BspMI BsrI BsrBI BsrDI BsrFI BsrGI BssHII Bst1107I BstBI BstEII BstNI BstUI BstXI BstYI Bsu36I ClaI Csp6I DdeI DpnI DpnII DraI DraIII DrdI EaeI EagI Eam1105I EarI Ecl136II Eco47III Eco57I EcoNI EcoO109I EcoRI EcoRV Esp3I Fnu4HI FokI FspI HaeII HaeIII HgaI HhaI HincII HindIII HinfI HinP1I HpaI HpaII HphI KasI KpnI MboI MboII MluI ...
Hi All, I am in trouble. I am developing an algorithm for restriction enzyme analysis but it is taking too long. The reason is because there are so many degenerate bases in the DNA sequence and thus it takes very long to analyze for all of them considering the possible combiantions they make. All the more the recognition sequence also have degenerate bases. Is there anybody out there to help me optimize the algorithm? Yes, there is. So thanking in advance to all those who respond. Ravi Gupta. Research Scholar DA University, M.P., India. -----------== Posted via Deja News, The Discussion Network ==---------- http://www.dejanews.com/ Search, Read, Discuss, or Start Your Own ...
This is an interface to a utility that determines the locations at which the selected enzyme is cutting the human sequence. If so specified, alternative enzymes that cut the sequence at the same location are reported. Enzyme: AatII AccI Acc65I AciI AflII AflIII AgeI AluI AlwI AlwNI ApaI ApaLI ApoI AscI AseI AvaI AvaII AvrII BamHI BanI BanII BbsI BbvI BcgI BcgI(second_site) BclI BfaI BglI BglII BpmI Bpu1102I BsaI BsaAI BsaBI BsaHI BsaJI BsaWI BsgI BsiEI BsiHKAI BsiWI BslI BsmI BsmAI BsmFI Bsp120I Bsp1286I BspDI BspEI BspHI BspMI BsrI BsrBI BsrDI BsrFI BsrGI BssHII Bst1107I BstBI BstEII BstNI BstUI BstXI BstYI Bsu36I ClaI Csp6I DdeI DpnI DpnII DraI DraIII DrdI EaeI EagI Eam1105I EarI Ecl136II Eco47III Eco57I EcoNI EcoO109I EcoRI EcoRV Esp3I Fnu4HI FokI FspI HaeII HaeIII HgaI HhaI HincII HindIII HinfI HinP1I HpaI HpaII HphI KasI KpnI MboI MboII MluI MnlI MscI MseI MslI MspI MspA1I MunI MwoI NaeI NarI NciI NcoI NdeI NgoMI NheI NlaIII NlaIV NotI NruI NsiI PacI PaeR7I PflMI PleI PmeI PmlI Ppu10I PpuMI ...
RECOMBINANT DNA RESEARCH, VOLUME 4: DOCUMENTS RELATING TO NIH GUIDELINES FOR RESEARCH INVOLVING RECOMBINANT DNA MOLECULES, AUGUST-DECEMBER ...
You have created a recombinant DNA molecule by ligating a gene to a plasmid vector. By mistake, your friend adds exonuelease enzyme to the tube containing the recombinant DNA. How will your experiment get affected as you…
Hi all: Im looking for a restriction enzyme that cuts human (eukaryotic) genomic DNA into smaller pieces than it does with bacterial (gram + actinomyces, GC-rich) chromosomal DNA. The purpose is to generate a bacterial chromosomal library from intracellularly grown bacteria (grown inside human macrophages) and to get rid of the human genomic DNA that may contaminate the bacterial DNA. If the human DNApieces are considerably smaller (lets say, below 10-15kb) than the bacterial ones (,25kb) the packaging of cosmids (Stratagene SuperCos) should exclude any DNA below 20kb (so they promise...). Does anybody know of such a restriction enzyme ? Or of a easily to deactivate DNAse ? Cheers, Markus schneema at cmgm.stanford.edu ...
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Synonyms for restriction fragment in Free Thesaurus. Antonyms for restriction fragment. 1 word related to restriction fragment: fragment. What are synonyms for restriction fragment?
Type II restriction enzymes are the molecular scissors that catalyse the double‐strand cleavage of deoxyribonucleic acid (DNA) at specific base sequences
Hi,. I am planning on buying the SAL1 restriction enzyme from new england biosystems. They dont offer DNA ligase in their kit. Im trying to cut portions of two separate genes with SAL 1 and link them together. Given that SAL 1 produces sticky ends,, do I need ligase to link my two gene segments together after restriction digest? If so, do I need some specific type of ligase? If some one could direct me to a company that offers SAL1 with applications/reagent requirements, I would be very much appreciative,. Thanks,. Dan. ...
Study Flashcards On restriction enzymes/plasmids at Cram.com. Quickly memorize the terms, phrases and much more. Cram.com makes it easy to get the grade you want!
The image shows the results of analyzing restriction enzyme digested PCR reactions from seven individuals using gel electrophoresis. Samples were loaded at the top of the gel shown below and run towards the bottom. The lanes on the gel are numbered at the top. Lane 1 contains what is called a "DNA ladder". This is a mix of DNA of known sizes that we purchase and run as a standard, or ruler, on the side of the gel. I have labeled the size of two of these "ladder rungs" or standards. You can use these standards to get an idea of the size of the bands in lanes 2-8. Each of these lanes represents one of seven individuals that have been tested ...
Blog on Oligo, Restriction Site, Avr II primer product: The Oligo, Restriction Site, Avr II n/a (Catalog #MBS634114) is a Primer and is intended for research pur...
The first four tracks correspond to the DNA strands listed above. The fifth and sixth tracks are 1k bp and 100 bp ladders. Tracks 7 and 8 correspond to lambda + HindIII and lambda + HindIII + EcoR1. Track 9 is dye and track 10 was a test track (done by one of the instructors). From this gel, you can see that we got a partial digest for KpnI and HindIII. Hernan redid the digest overnight so that we would have better product to work with. Step 3: PCR amplification (Mon morning) - the next thing we did was to purify our sample. We started with the DNA from the gel and used a kit to purify it (remove the agarose, etc). After the PCR (?), we use a nanospectorphotometer in the Phillips lab to find the concentration of DNA (14.7 ng/ul). Step 4: ligation (Mon morning) - once we had our purified DNA, it was time to ligate it to the lacZ that had been pulled out of the pZE21 plasmid. We used three different ratios of vector (pZS25) to insert (lacZ) - 1:3, 1:1 and 3:1. We also ran a 0 insert control. Once ...
The first four tracks correspond to the DNA strands listed above. The fifth and sixth tracks are 1k bp and 100 bp ladders. Tracks 7 and 8 correspond to lambda + HindIII and lambda + HindIII + EcoR1. Track 9 is dye and track 10 was a test track (done by one of the instructors). From this gel, you can see that we got a partial digest for KpnI and HindIII. Hernan redid the digest overnight so that we would have better product to work with. Step 3: PCR amplification (Mon morning) - the next thing we did was to purify our sample. We started with the DNA from the gel and used a kit to purify it (remove the agarose, etc). After the PCR (?), we use a nanospectorphotometer in the Phillips lab to find the concentration of DNA (14.7 ng/ul). Step 4: ligation (Mon morning) - once we had our purified DNA, it was time to ligate it to the lacZ that had been pulled out of the pZE21 plasmid. We used three different ratios of vector (pZS25) to insert (lacZ) - 1:3, 1:1 and 3:1. We also ran a 0 insert control. Once ...
the amount of enzyme for the quantity of plasmid used is quite high. I recommend either increasing the volume of the digest (~50ul) or using 0.5ul of EcoRI.. ...