cloning of polymerase chain reaction pcr amplified

... Alignment of nucleotide sequence of the 5’- and 3’-termini of mustard GADs 156 32 Prediction of the secondary structure of C-terminal of mGADs with PSIPRED software 158 33 Hydropathy profile of cDNA-derived ... reverse transcription PCR (RT -PCR) 71 3.11 Cloning of full-length cDNA by RACE 72 3.11.1 5’-RACE 72 3.11.2 3’-RACE 74 3.11.3 Generation of full-length cDNA sequences 74 Cloning of the BjDAO promoter ... he also gives me professional support, help and inspiration i Table of Contents Page Acknowledgements i Table of Contents ii List of Abbreviations x List of Figures xiii List of Tables xvii Summary...

Ngày tải lên: 12/09/2015, 10:19

... specific DNA segments • Polymerase Chain Reaction • Requires knowledge of the DNA sequence in the region of interest • As more sequence information becomes available, the uses of PCR expand • With ... Denature Anneal primers Synthesize new DNA with polymerase Polymerase chain reaction, cycle Cycle Denature Anneal primers Synthesize new DNA with polymerase PCR, cycle Cycle (focus on DNA segments bounded ... of desired product PCR: make large amounts of a particular sequence • The number of molecules of the DNA fragment between the primers increases about 2-fold with each cycle • For n = number of...

Ngày tải lên: 13/03/2014, 18:39

... http://www.biomedcentral.com/1471-2229/7/66 Figure profile of the assembly of cassava ESTs Cluster Cluster profile of the assembly of cassava ESTs The graph presents the number of clones per assembled scaffold; ... transcripts of a total of 7796 distinct genes, with alternative transcripts of about 26% of them To find the number of new transcripts captured in this library, relative to the number of expressed ... best hit of each sequence of cassava in each of the other databases and the best hit of each sequence of the other species in cassava We thus built a table representing the network of best hits...

Ngày tải lên: 12/08/2014, 05:20

... detection of C pneumoniae in human specimens are polymerase chain reaction (PCR) , in-situ hybridization method and enzyme immunoassay protocols (12, 13) PCR- based approach usually targets parts of chlamydial ... µl of DNA was added to reaction mixture containing 1x PCR buffer (Silex, Moscow, RF) containing 10 mM Tris-HCl, pH 8,3 , 2.5 mM MgCl2, 15 pmol of each primer, 200 µM of each of dNTPs and U of ... round of PCR, µl of the first-round product was mixed with 23 µl of amplification mixture containing primers for iCPC and iCPD and amplified using following cycling conditions: 35 cycles of denaturation...

Ngày tải lên: 26/10/2012, 08:57

... cis-prenyltransferases, was termed LT600 3¢- and 5¢-RACE reaction and cloning of Hevea cis-prenyltransferase cDNA Cloning of the full-length cDNA of Hevea cis-prenyltransferase, used poly(A)+ RNA from ... IPTG; S2, supernatant of the homogenate of E coli BL21(DE3)/pETHRT2 with IPTG; PC, pellet of the homogenate of E coli BL21(DE3)/pET32b without IPTG; P1, pellet of the homogenate of E coli BL21(DE3)/pETHRT1 ... Conversely, the reaction of WBP or HRT2 alone gave a small spot at the origin of the TLC plate In order to determine the molecular masses of the rubber products from the reaction of HRT2 coincubated...

Ngày tải lên: 07/03/2014, 21:20

... amounts of RNA loaded per lane The sizes of an RNA marker are indicated the cloning of the 5¢ and 3¢ end of the clone, respectively Analysis of the nucleotide sequences of the newly obtained PCR- fragments ... peroxidase eluted at Fig Determination of the pH-optimum of the foot-specific peroxi320–360 mM NaCl (Fig 3B), the yield of activity was 66% dase Equal amounts of an extract of foot pieces were reacted with ... incubation time of 30 min, the reaction bound to the column and was eluted by decreasing the ionic was stopped by the addition of 10 lL of 100 mM NaN3, and the strength Hydroxyapatite often resolves...

Ngày tải lên: 08/03/2014, 10:20

... sites They were then inserted at the multicloning site of pSHLeu in frame For the disruption of MRP4, a 1.8-Kb DNA fragment containing the MRP4 gene was PCR amplified and cloned into pUC119 Then, ... Disruption of the mrp gene in each of them was confirmed by PCR (data not shown) Growth media, culture conditions and genetic manipulations are essentially as described [20] For the preparation of mitochondria, ... medium and showed slow growth on YPD as in the case of disruptants of most other mrp genes In the case of MRPS8, it was indicated that the addition of a short peptide to the C-terminus caused poor...

Ngày tải lên: 17/03/2014, 10:20

... template by polymerase chain reaction using primers BPL-for and BPL-rev; and BCCPD67 and BCCP-rev, respectively The PCR products were cloned into plasmid pCR2 .1 (Invitrogen) using standard TOPO cloning ... BCCPD67 K117L and the plasmid pCR2 .1/BccpD67 was used as the PCR template The mutant gene PCR product was cloned into pCR2 .1 and the resulting plasmid was named pCR2 .1/BccpD67 K117L To express ... apo-BCCPD67 The reaction was initiated by the addition of purified BPL to a final concentration of lM, and incubated at 70 °C for 30 The reaction was terminated by the addition of ice-cold trichloroacetic...

Ngày tải lên: 31/03/2014, 07:20

... three-dimensional structures of certain domains of nitric oxide synthase isoforms are known [53±57], we are not aware of the three-dimensional structure of the calmodulin binding sites of any isoform being reported ... 82±101 of caveolin-1 in the form of glutathione S-transferase fusion proteins have been shown to attenuate the activities of all three isoforms of NOS [62] A novel 10-kDa protein inhibitor of nNOS, ... molecule discovery program of the Australian Institute of Marine Science It effectively inhibited nNOS with an IC50 value of 16.2 lM with a slope of 1.5 This is the ®rst instance of an amphibian peptide...

Ngày tải lên: 31/03/2014, 15:20

... the roof If additional roof deflection is mm or less, the isolator should be selected for 10 times the additional roof deflection If additional roof deflection is over mm, supplemental roof stiffening ... class of equipment Certain spaces may dictate higher levels of isolation For example, bar joist roofs may require a static deflection of 38 mm over factories, but 64 mm over commercial office ... class of equipment Certain spaces may dictate higher levels of isolation For example, bar joist roofs may require a static deflection of 38 mm over factories, but 64 mm over commercial office...

Ngày tải lên: 31/03/2014, 18:20

... (5′-dCTCTGCGGCCGCT17-3′) 20 PCR reaction mix: µL of 10X PCR buffer, µL of Taq DNA polymerase, 2.5 µL each of NotI-dC and SalI-dT primer, µL of 25 mM MgCl2, µL of 10 mM dNTPs, 35 µL of water; prepare just ... prepared: µL of 10X PCR buffer (provided with the polymerase mixture), 1.5 µL of dNTP mix (10 mM of each), µL of diluted cDNA template, polymerase mixture sufficient for 60 µL of PCR (see manufacturer’s ... amplified by polymerase chain reaction (PCR) Since the initial publication in 1988 (1), RACE has greatly facilitated the cloning of new genes Currently, RACE remains the most effective method of cloning...

Ngày tải lên: 11/04/2014, 01:04

... 20% The direct costs of the production of m3 of raw wood assortments were derived by means of the product of SH/m3 with the wage schedule and coefficient of recalculation of labour costs to direct ... difference of gross yield and own costs of logging Value yield tables of poplar clones were constructed by the method of simulation from the models of assortment yield tables, prices of the assortments ... fusion of models of yield tables and stand assortment tables, models of quality and models of damage to stands Models of yield tables (Petráš, Mecko 2001, 2005) determine the development of mean...

Ngày tải lên: 07/08/2014, 03:22

... Spearman’s coefficient of rank correlation (rs) This test was used to evaluate the stability of the degree of infection within the particular clones of black poplar in the years of observation, the ... from clones 7, 107, 110, 152 The stability of the intensity of infection of black poplar clones by the rust (the rank of clones according to the intensity of infection) between the particular years ... the time frame of the main incidence of the rust Stem diameter and the main part of plant height were realized before the onset of the pathogen incidence while the production of storage compounds...

Ngày tải lên: 07/08/2014, 10:21

... diameter of stand (cm) Fig Proportions of the quality classes of logs of clone Robusta with zero and 40% damage of stems of quality class A Fig Proportions of the quality classes of logs of clone ... classes of logs of clone Robusta with zero and 40% damage of stems of quality class B Fig Proportions of the quality classes of logs of clone I-214 with zero and 40% damage of stems of quality ... 10 Damage of stands 0% 90 VI VI 14 18 22 26 30 34 38 42 46 50 54 58 62 Mean diameter of stand (cm) Proportion of quality classes of the logs (%) 0% 90 Proportion of quality classes of the logs...

Ngày tải lên: 07/08/2014, 10:21

... the PCR result of two isolates with primers specific for subgroup J ALVs The isolates were positive in PCR, however, the PCR product size of one isolate(KOALPET) was was smaller than that of ADOL-Hc1 ... was interesting that the PCR product size of a Korean isolate was somewhat different from ADOL-Hc1 The PCR product size of one isolate, KOAL-PET, was smaller than that of ADOL-Hc1 The primer sequences ... isolate(KOALPET) was was smaller than that of ADOL-Hc1 Fig PCR reaction of Korean isolates with subgroup J specific primer pair S1/S2 Lane 1, MW marker of 100bp multiples(Gibco BRL); lane 2, KOAL-PET;...

Ngày tải lên: 07/08/2014, 15:20

... In vitro cholesterol-lowering test: Among isolated 100 strains of LAB, of Streptococcus strains, 11 of Lactobacillus strains and of Bifidobacterium strains were selected as provisional probiotic ... presence of bile salt These results showed that many LAB have a great cholesterol-lowering activity in the MRS broth regardless to the presence of bile salt (0.3% oxgall) (Table 2) Identification of ... vande Woestyne M, Verstraete W In vitro study of bile-salt hydrolase (BSH) activity of BSH isogenic Lactobacillus plantarum 80 strains and estimation of cholesterol lowering through enhanced BSH...

Ngày tải lên: 07/08/2014, 18:20

... this article, we report the micropropagation of late-flushing Q robur from acorns of selected trees To our knowledge, results of ex vitro rooting of micropropagated oak are presented here for ... for and then rinsed in sterile water times for In each case, half of the embryos of the 11 mother trees were placed on solid Gresshoff and Doy (GD) medium (1972) and woody plant medium (WPM) (Lloyd ... the individual clones The influence of the genetic background of the mother tree is demonstrated by differences found in plant survival by the mean number of surviving plants/clone (table I)...

Ngày tải lên: 08/08/2014, 19:21

... development of each individual within a particular stand This development may be influenced by other factors, eg, the availability of limiting resources This may be the origin of size hierarchies of individuals ... 1990) and at the same time in a growing size inequality The objectives of this paper are twofold: (1) to characterize a number of poplar cultivars by some statistical parameters (ie size inequality); ... and the position of individual trees MATERIALS AND METHODS Study areas A short rotation intensively cultured (SRIC) plantation of poplar (Populus sp) was grown at the location of Afsnee (51° °...

Ngày tải lên: 08/08/2014, 19:21

... characteristics was collected on representative trees of each of the clones Number of branches, branch length and diameter, and the angles of origin and termination were determined for both proleptic ... measured Dry weight (DW) of proleptic and sylleptic branches, current terminal and the remainder of the stem were determined, as well as DW and leaf area (LA) of leaves of current terminal, proleptic ... the middle of the growing season, 35-40% of the total LA were carried on sylleptic branches in hybrid clones 44-136 and 11-11, respectively The LA on the current terminal of each of the clones...

Ngày tải lên: 09/08/2014, 02:21

... Tab I) 2.2 PCR conditions PCR reactions were performed on an MJ Research PTC-100 thermocycler in 15 àL reaction volumes with 1ì Mg2+ free buffer, 0.125 mM dNTP, 1.5 mM MgCl2 , 0.5 àM of each primer ... presence of the gene of interest was conrmed by PCR on the BAC DNA For the three BAC clones identied using PCR systems and BAC for the Texas marker genes 197 Figure Metaphase spread of bovine ... Hors-Cayla [9] and is composed of a total of 38 hamster-bovine cell lines A more complete description of the panel is given in Laurent et al [11] A correlation coefcient of 0.69 was used as the threshold...

Ngày tải lên: 09/08/2014, 18:21