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T1 transgenic tobacco plants
T1 transgenic tobacco plants

... 3.2-A). ZmDEF1 gene cloned with these steps: pBT attached to the vector; DH5α transformed into E. coli by heat shock; colony PCR containing target gene to select line recombinant using primers M13_F / M13_R (Figure 3.2-C). Before sequencing, gene recombinant plasmid ZmDEF1 was checked by BamHI cut t ...
Careful risk assessment needed to evaluate transgenic fish
Careful risk assessment needed to evaluate transgenic fish

... fish have been developed for applications such as the production of human therapeutics, experimental models for biological research, environmental monitoring, ornamental fish and aquacultural production. Ironically, in addition to being the taxonomic group with the most transgenic species, aquatic org ...
Genetic Transformation of Poinsettia (Euphórbia
Genetic Transformation of Poinsettia (Euphórbia

... Ås, 13th of August 2015 ...
Rh phenotype prediction by DNA typing and its
Rh phenotype prediction by DNA typing and its

... group genotyping applications. The discrimination of RHD homozygotes from RHD heterozygotes is hampered (Cossu et al., 1996) because the most prevalent RHD-negative allele cannot yet be detected specifically. For the time being, we prefer therefore the term DNA typing rather than genotyping, when an ...
Myriad--Ambry -- Final Version of Ambry Preliminary Injunction
Myriad--Ambry -- Final Version of Ambry Preliminary Injunction

... (the “template strand”) is actually transcribed into mRNA and ultimately used to produce a protein, while the other strand is, for purposes of that gene, simply a space-filling molecule to provide the other side of the double helix. This space-filling strand is sometimes referred to as the “coding s ...
Chondroitinase ABC I from Proteus vulgaris: cloning, recombinant
Chondroitinase ABC I from Proteus vulgaris: cloning, recombinant

... matrix) and at the cell surface as constituents of proteoglycans. These sugars, apart from having important structural roles in the ECM, are also fundamental modulators of many biological processes, such as development, cell proliferation, signalling and inflammation [1–3]. The great chemical hetero ...
TRIzol Reagent
TRIzol Reagent

... BCP (1-bromo-2 chloropropane) can be used in the place of chloroform. BCP has a higher density. Because of its higher density, the interface is more compact and therefore the aqueous layer is more easily recovered. (see Chomczynski, P. and Mackey, K. (1995) Anal. Biochem. 225, 163-164). The quantity ...
Stage-specific expressions of four different ribonuclease H genes in
Stage-specific expressions of four different ribonuclease H genes in

Chromosome - World of Teaching
Chromosome - World of Teaching

... The DNA of eukaryotic cell is tightly bound to small basic proteins (histones) that package the DNA in an orderly way in the cell nucleus. This task is substantial (necessary), given the DNA content of most eukaryotes For e.g., the total extended length of DNA in a human cell is nearly 2 m, but this ...
Getting Started With Gel Electrophoresis
Getting Started With Gel Electrophoresis

... The composition of this buffer is designed to keep the pH slightly alkaline. This ensures that the DNA molecules stay in their deprotonated form which includes a negative charge on each phosphate group. Without that negative charge, DNA molecules would not be moved by an electric field and electroph ...
PPT - Altogen Biosystems
PPT - Altogen Biosystems

... 3. Prepare transfection complexes by mixing 40 µl of serum free medium, 5.5 µl of transfection reagent, and • 750 ng DNA (or mRNA), or • 30 nM - 50 nM of siRNA (or microRNA) *Referred to a final volume including growth medium 4. Incubate transfection complexes at RT for 15 - 30 minutes 5. Optional: ...
Action
Action

... Shanghai Jiao Tong University ...
Harvey ras (H-ras) Point Mutations Are Induced by 4
Harvey ras (H-ras) Point Mutations Are Induced by 4

... stained cryosections and dissected (Fig. 1). Genomic DNA was iso lated from the lesion and amplified by nested PCR. RFLP Analysis. 4NQO acting as a DNA alkylating agent should result in 0 to A transversion (4,5). Of the codons previously impli cated in H-ras activation, codon 12 has two available G ...
transposon
transposon

... is given the prefix IS, followed by a number that identifies the type.  The IS elements are normal constituents of bacterial chromosomes and plasmids. To describe an insertion into a particular site, a double colon is used; so λ::IS1 describes an IS1 element inserted into phage lambda.  The IS ele ...
Regulation of Bacterial Conjugation in Microaerobiosis by
Regulation of Bacterial Conjugation in Microaerobiosis by

... 39 and 59 CCG GAA TTC CGC AAG CTG AGA TAA ACA GC 39. The resulting fragment was purified with the Wizard SV Gel and PCR Clean-Up System (Promega, Madison, WI). After digestion with BamHI and EcoRI the amplified fragment was cloned onto pGEX4T-1 (GE Healthcare, Little Chalfont, UK) to obtain a fusion ...
tutorial - ppuigbo
tutorial - ppuigbo

... tab-delimited format for copying and pasting them into a ...
The physics behind the larger scale organization of DNA in eukaryotes
The physics behind the larger scale organization of DNA in eukaryotes

... experiment a 167 bp repeat length was also probed, resulting in a 21.3 nm wide fiber with an NLD of 0.56 nucleosome nm−1 [19]. This might point to a 3-ribbon structure. The small discrepancy can be attributed to the approximations involved in the model that start to matter at such small fiber diamet ...
SNP-Based Mapping of Crossover Recombination in
SNP-Based Mapping of Crossover Recombination in

... tribution along chromosomes. In addition, some morphological markers can have effects on the viability ofhomozygotes. An alter­ native approach, first pioneered by Wicks et al. ( 1) for gene map­ ping, involves the use of mapped sequence differences between tvvo laboratory strains of C. elegans. The ...
Analysis of Cross Sequence Similarities for Multiple - PolyU
Analysis of Cross Sequence Similarities for Multiple - PolyU

... and complementary palindromes repeats are encoded by the LZ-based algorithm, whereas short subsequences are compressed using CTW. Although it obtains good compression ratio, its execution time is too high for long sequences. DNACompress (Chen et al., 2002) consists of two parts. All approximate repe ...
Get  - Wiley Online Library
Get - Wiley Online Library

... are planar. Although several tautomeric forms exist in principle, the keto and amino forms predominate (Figure 2). Typically, the enol and imino forms are no more than 0.1% of the total, although this amount can change dramatically if modified bases are considered or if base pairs are formed. Details ...
PPT - Altogen Biosystems
PPT - Altogen Biosystems

... Products > 4T1 Transfection Reagent (Breast Cancer Cells, CRL-2539) ...
Article Mitochondrial DNA turnover occurs during preimplantation
Article Mitochondrial DNA turnover occurs during preimplantation

... Figure 1. MtDNA turnover during mouse preimplantation development is sensitive to environmental factors. Full grown oocytes (GV) were isolated from ovaries, unfertilized eggs (UF) were collected 12 h after HCG injection, and fertilized eggs (FE), 2-, 4-, 8-cell and blastocyst stage embryos were coll ...
(lip) that - Repositories
(lip) that - Repositories

... Previous studies with enteric bacteria have identified a single genetic locus (lip) that is presumed to encode a lipoic acid biosynthetic enzyme (Herbert and Guest, 1968). The lip locus is situated at 14 minutes on the Salmonella typhimurium genetic map and is cotransducible with the leuS and nadD g ...
Analysis of acid-induced asr gene promoter of Enterobacteriaceae
Analysis of acid-induced asr gene promoter of Enterobacteriaceae

... Deletion analysis of asr promoter region upstream the proposed pho box. In order to identify potential cis-regulatory sites in the asr promoter, deletion analysis of the DNA region upstream the –40 position was performed. The consecutive promoter deletions (p∆70, p∆37, p∆20, p∆21, p∆10, p∆4, p∆1 in ...
Development of Co-Dominant Amplified Polymorphic Sequence
Development of Co-Dominant Amplified Polymorphic Sequence

... The original cross to generate the RILs utilized upland japonica cv. Moroberekan and susceptible indica cv. CO39. The intersubspecies cross was predicted to possess sufficiently high rates of polymorphism to efficiently generate CAPS markers. Eight CAPS markers, including S12886, were found at the P ...
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Molecular cloning



Molecular cloning is a set of experimental methods in molecular biology that are used to assemble recombinant DNA molecules and to direct their replication within host organisms. The use of the word cloning refers to the fact that the method involves the replication of one molecule to produce a population of cells with identical DNA molecules. Molecular cloning generally uses DNA sequences from two different organisms: the species that is the source of the DNA to be cloned, and the species that will serve as the living host for replication of the recombinant DNA. Molecular cloning methods are central to many contemporary areas of modern biology and medicine.In a conventional molecular cloning experiment, the DNA to be cloned is obtained from an organism of interest, then treated with enzymes in the test tube to generate smaller DNA fragments. Subsequently, these fragments are then combined with vector DNA to generate recombinant DNA molecules. The recombinant DNA is then introduced into a host organism (typically an easy-to-grow, benign, laboratory strain of E. coli bacteria). This will generate a population of organisms in which recombinant DNA molecules are replicated along with the host DNA. Because they contain foreign DNA fragments, these are transgenic or genetically modified microorganisms (GMO). This process takes advantage of the fact that a single bacterial cell can be induced to take up and replicate a single recombinant DNA molecule. This single cell can then be expanded exponentially to generate a large amount of bacteria, each of which contain copies of the original recombinant molecule. Thus, both the resulting bacterial population, and the recombinant DNA molecule, are commonly referred to as ""clones"". Strictly speaking, recombinant DNA refers to DNA molecules, while molecular cloning refers to the experimental methods used to assemble them.
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