cloning dna sequencing and sequence analysis

... to allow a large number of clones to be sequenced Cloning, DNA sequencing and sequence analysis The 16S rDNA amplicons were inserted into a vector using a cloning kit (pGEM-T vector; Promega, Madison, ... some bacterial DNA sequences not previously characterized by rDNA sequencing since they exhibit less than 97% similarity to known database sequences For example, DNA from the candidate division ... procedures, and performed clinical and rheumatological data analyses AZ, CB and EC conducted assessment of Chlamydia trachomatis serology and DNA extraction BJ and JS participated in the design and coordination...

Ngày tải lên: 09/08/2014, 10:23

... S-transferase (GST)-L1 and GST-L1(DL2) fusion proteins, was subjected to SDS/PAGE and transferred to a PVDF membrane The 28-kDa band on a membrane was subjected to amino-acid sequence analysis Cloning of ... conditions and two bands of 33 and 40 kDa under nonreducing conditions 2-ME, 2-mercaptoethanol (B) The N-terminal amino-acid sequences of three bands The 28-kDa protein had a single amino-acid sequence ... formaldehyde, and RNA bands were transferred to a HybondN+ nylon membrane (Amersham) Stringency used for hybridization and washing, and the probe were the same as those, used in cloning After...

Ngày tải lên: 17/03/2014, 11:20

... Nucleotide sequences were aligned and phylogenetic analysis was performed using the Multiple sequence alignment program, AlinX (Vector NTI Suite version 5.2.1.3.; InforMax, USA) Results Genomic DNAs ... province (Table 1) The 396 bp PCR product o o o o o o o o o o o Cloning and sequence analysis PCR amplicons were purified using a GFX PCR DNA Purification Kit (Amersham, UK) according to the manufacturer’s ... and adult ticks from cattle, horses, goats, dogs, cats, hedgehogs and wild rodents from Korean provinces [18] The ticks were identified and categorized with respect to developmental stages, and...

Ngày tải lên: 07/08/2014, 18:21

... the other hand, despite being costly or technically demanding, LiPA and direct DNA sequencing yielded accurate identification of all genotype A and genotype D and mixed genotype A and D strains ... SA and WAN designed the study and FH collected the specimens MMA carried out the DNA Extraction, INNO-LiPA genotyping assay, Direct DNA Sequencing, PCR-RFLP studies, sequence alignments and analysis ... DNA was isolated from 10 independent clones of Escherichia coli by a modified alkaline lysis procedure [12] and sequenced by using quick start cycle DNA sequencing kit (Beckman-Coulter) The DNA...

Ngày tải lên: 12/08/2014, 04:20

... products and allows the DNA sequence to be read Either primers or ddNTPs can be fluorescently labeled allowing detection in an automated DNA sequencer and Bst DNA polymerase I (8) (BioRad) The DNA ... allow linear accumulation of one strand of DNA suitable for subsequent DNA sequencing Analysis, sequencing and in vitro expression of PCR products 99 Template DNA PCR with one biotinylated primer ... shows an example of DNA sequence from purified DNA with AmpliTaqFS and fluorescent-labeled ddNTPs with analysis on an ABI 373 DNA sequencer allows simple separation of the solid and solution phases...

Ngày tải lên: 25/10/2013, 22:20

... humans, and then subsequently in mouse, rat and other mammalian counterparts [26–28] In the present work we isolated and characterized a carp cDNA sequence that is homologous to the DNA sequence ... human and pufferfish (torafugu and spotted green pufferfish) IL-10 sequences Pufferfish and carp IL-10 genes, clustered together and distant from IL-20 and IL24, as recently determined from analysis ... Phylogenetic Analysis Using Parsimony ( *and Other Methods), 4th edn Sinauer Associates, Sunderland, Massachussetts 24 Laing, K.J., Holland, J., Bonilla, S., Cunningham, C & Secombes, C.J (2001) Cloning and...

Ngày tải lên: 20/02/2014, 02:21

... (Stratagene, La Jolla, CA, USA) and subjected to DNA sequence analysis [62] Rapid amplification of cDNA ends (RACE) Analysis by 5¢- and 3¢-RACE was performed using a SMARTTM RACE cDNA amplification kit (Clontech ... thereof Sense and antisense primers based on the resulting cDNA sequences were selected and used to amplify full-length cDNAs by RACE PCR, resulting in 11 full-length and two partial cDNA clones ... G13 variants, G19 and G19 h all contained an R & R consensus sequence, and they exhibited significant sequence similarity (G19 and G13, 58% identity; G19 h and G13, 48%; G19 and G19 h, 64%) A...

Ngày tải lên: 07/03/2014, 21:20

... encompassing the conserved sequence boxes I-III (CSB), tRNA genes, L and H strand promoters (LSP and HSP), L and H strand transcription start sites (LSS and HSS) The L-strand sequence and position of the ... OL and OH represent the origin of L and H strand replication, respectively LSS, L strand start site; HSS, H strand start site The map positions of the rRNA, tRNAs and mRNAs (ND1 to ND6, A6 and ... piperidine and analysis of cleaved DNA strands on a sequencing gel, were carried out as described before [26,27] UV induced protein -DNA cross-linking A photoreactive 32P-labelled D-TERM DNA probe...

Ngày tải lên: 08/03/2014, 08:20

... the sequenced position This sequencing results to the sequences of quantities and lengths comparable to illumine sequencing 10 DNA Sequencing – Methods and Applications 6.6 DNA nanoball sequencing ... MgCl2 for standard and CleanAmpTM), B4GN4 (90% - a mixture of the 1.5 and mM MgCl2 for standard dNTPs and 100% - mM MgCl2 for standard and CleanAmpTM), GNAQ (100% - mM MgCl2 for standard and CleanAmpTM), ... speed of sequencing attained with modern DNA sequencing technology has been instrumental in the sequencing of the human genome, in the human genome project Fig DNA Sequence Trace DNA can be sequenced...

Ngày tải lên: 08/03/2014, 19:20

... sites After binding and hydroxyl radical attack, free (F) and bound (B) DNA were separated and analysed The left and right panels in (A) and (B) represent the top and bottom strands of the binding ... Experimental procedures Cloning, expression and purification of recombinant proteins HAKN1 homeodomain and C-terminal sequences were amplified and cloned in-frame into the EcoRI and PstI sites of the ... the sequence GTNAC, postulated to be important for the binding of the tobacco protein NTH15 to DNA [24], provided that N is G or C Analysis of DNA binding by hydroxyl radical footprinting and...

Ngày tải lên: 23/03/2014, 13:20

... sequences (254 and 106 bp) between the NEGU and NEG-D elements The distance between the NEG-U and NEG-D elements was 253 and 106 bp in D-254 and D-106, respectively The right-hand panel shows ... in F-560 and M-B indicates that both the NEG-U and NEG-D elements were mutated in F-560 (B) The lefthand panel shows F-560 and different mutant constructs harboring shorter intervening sequences ... deletion analysis, mutation analysis, as well as decoy assays, and found that the NEG-U, NEG-D and POS-D elements play important roles in regulation of the Pokemon promoter; this helps us understand...

Ngày tải lên: 30/03/2014, 04:20

... Results Cloning and sequencing of the StP gene Using the PCR primers p1 and p2, a 710-bp fragment was amplified from chromosomal C callunae DNA, blunt-end cloned into pUC 18, and sequenced The sequence ... elsewhere [11,12] Restriction endonucleases, T 4DNA ligase and Taq DNA polymerase were obtained from Promega Pfu DNA polymerase, alkaline phosphatase, RNase, and positively charged nylon membranes were ... primary structure of StP and report here the cloning, sequencing and heterologous expression in Escherichia coli of the gene encoding this enzyme In an effort to identify sequence changes relevant...

Ngày tải lên: 31/03/2014, 01:20

... the Macintosh sequence analysis program DNA Strider ; ### from DNA Strider ;-) ; DNA sequence seq1, 16 bases, 2688 checksum ; agctagctagctagct // ; ### from DNA Strider ;-) ; DNA sequence seq2, ... in the sequence, one to sequence more of the same strand (purple) and a second (turquoise) that hybridizes to the complementary strand and produces a sequence running backward on this strand, thus ... sequence entry format 32 s CHAPTER Figure 2.8 FASTA sequence entry format sequence; (2) the sequence in standard one-letter symbols; and (3) an optional “*” which indicates end of sequence and...

Ngày tải lên: 08/04/2014, 12:44

... investigated and utilized in model-based algorithms and extended gradient-based algorithms Sudhir et al [13] and Clifford and Nasrabadi [14] used Markov random fields to combine the stereo and motion ... technique Horizontal and vertical coordinates (x, y) can be measured because horizontal and vertical displacements Δx and Δy between the imagescan be measured by the motion analysis, and the initial ... system, two images (left and right) sequences can be simultaneously acquired This enables acquisition of a pair of motion fields by analyzing the left and right image sequences Since true motion...

Ngày tải lên: 21/06/2014, 22:20

... explained in [36] Spectral and wavelet analyses of symbolic sequences are explained and applied to DNA sequences, and results are presented for “pseudo DNA sequences and E Coli DNA Concepts from digital ... SEQUENCE ANALYSIS A primary objective of DNA sequence analysis is to automatically interpret DNA sequences and provide the location and function of protein coding regions Methods to locate genes, and ... DNA sequences are discussed in Section 4.2 4.2 Proposed AR model-based DNA sequence analysis The AR modeling of a DNA sequence is done by first mapping the sequence into the numerical domain and...

Ngày tải lên: 23/06/2014, 01:20

... canine LINE-1 and was followed by a 5bp deletion; DNA sequences of the TVT-LINE showed a 98 % and 63 % homology to canine LINE-1 and human LINE-1 sequences, respectively Sequence Analysis of Canine ... from the gel, were determined for DNA sequences directly, and the DNA sequences of the TVT-LINE were aligned with those of the canine ORF2 region (Fig 2) The DNA sequences of the inserts from three ... Synthetic DNA primers were used for the PCR, RT-PCR, LA-PCR and DNA sequencing The DNA sequence data were analyzed by GENETYX-MAX software (Software Development Co Ltd, Japan) for multiple sequence...

Ngày tải lên: 07/08/2014, 15:20

... methylation-filtered and 11% of high-CoT sequences were similar to known plant expressed sequence tags (ESTs), whereas only 4% of sequences from random libraries were similar Palmer and coworkers [9] ... Tibbitts DC, Wessler SR, Paterson AH: Integration of Cot analysis, DNA cloning, and high-throughput sequencing facilitates genome characterization and gene discovery Genome Res 2002, 12:795-807 Palmer ... genome sequencing project Plant Physiol 2001, 127:1572-1578 Lander ES, Linton LM, Birren B, Nusbaum C, Zody MC, Baldwin J, Devon K, Dewar K, Doyle M, FitzHugh W, et al.: Initial sequencing and analysis...

Ngày tải lên: 09/08/2014, 20:20

... Raw sequence trace data were processed to screen out vector and linking sequence, to remove low-quality sequence, and to trim poly(dA) tails using an in-house software solution The resulting sequences ... S: Identification of potential vaccine and drug target candidates by expressed sequence tag analysis and immunoscreening of Onchocerca volvulus larval cDNA libraries Infect Immun 2000, 68:3491-3501 ... these, three showed database homologies from 3' sequence and were reclassified as conserved, and two showed no poly(A) tail and were excluded from further analysis as presumed internal fragments The...

Ngày tải lên: 09/08/2014, 20:20