extraction cdna synthesis and qrt pcr

... http://www.biomedcentral.com/content/supplementary/1471-2229-10-2S1.DOC ] Additional file 2: Primers used for mapping, RT -PCR and qRT- PCR displaying primers used for mapping, RT -PCR and qRT- PCR Click here for file [ http://www.biomedcentral.com/content/supplementary/1471-2229-10-2S2.DOC ... base, and did preliminary molecular analysis for qRT- PCR; CR, performed in silico search of maize ZEPs, primer design and sequencing for confirming in silico analyses, applying to qRT- PCR and mapping ... [34] RT -PCR and qRT- PCR Gene expression patterns using RT -PCR were determined as previously described [54] In these end point analyses, the cycle number was adjusted to yield nonsaturated bands...

Ngày tải lên: 12/08/2014, 03:21

... the Cc RNase cDNA The selected hybrids were amplified by PCR and cloned into the pCR 2.1 vector Twenty-four cDNA clones were selected and verified by restriction endonuclease mapping and DNA sequence ... Genomic structure and expression analysis of the Cc RNase epidermis and intestine of 6-day-old larvae and adult heads and ovaries, was blotted and hybridized using the Cc RNase cDNA as a probe ... were then amplified by PCR using the LD primer (5¢-AAGCAGTGGTAACAACGCAGAGT-3¢), which anneals at the 5¢ and 3¢ ends of the cDNA strands The PCR products were cloned into the pCR 2.1 cloning vector...

Ngày tải lên: 23/03/2014, 06:20

... migration profiles of core+1 ⁄ F–myc and core+1 ⁄ S–myc proteins, at  22 and 13 kDa, respectively, are indicated by the open and filled arrowheads The arrows show core and actin The migration positions ... HCV-1 core ⁄ core+1 sequence, and the primer pair C53–C203 (Table 2) First, the C53–C203 PCR product was digested with EcoRI and inserted into the EcoRI cloning site of pcDNA3.1(–) ⁄ Myc-His B (Invitrogen, ... codons 8–11 myc myc myc – 342–825 pHPI-1494 (pcDNA3.1(–) ⁄ Myc-His) pHPI-1507 (pcDNA3.1(–) ⁄ Myc-His) pHPI-1495 (pcDNA3.1(–) ⁄ Myc-His) pHPI-1705 (pcDNA3.1(–) ⁄ Myc-His) Mutation Tag molecule Length...

Ngày tải lên: 30/03/2014, 03:20

... start to walk and refuse to rest They just want to walk but have no idea where they are walking to Some patients clap their hands on and on, others fold and refold laundry over and over They ... showed later that mean p25/p35 ratios were about 1.7 and for control and AD brains respectively (Tseng et al., 2002) One the other hand, Tandon and his colleagues pointed out that no elevation of ... beta-amyloid deposits and other obvious pathological lesions Later, cross-linked and argyrophilic precipitates start to occur The distal segments of the dendrites turn to twisted and dilated and probably...

Ngày tải lên: 05/10/2015, 22:32

... start to walk and refuse to rest They just want to walk but have no idea where they are walking to Some patients clap their hands on and on, others fold and refold laundry over and over They ... showed later that mean p25/p35 ratios were about 1.7 and for control and AD brains respectively (Tseng et al., 2002) One the other hand, Tandon and his colleagues pointed out that no elevation of ... beta-amyloid deposits and other obvious pathological lesions Later, cross-linked and argyrophilic precipitates start to occur The distal segments of the dendrites turn to twisted and dilated and probably...

Ngày tải lên: 06/10/2015, 20:35

... comprises at least two genes ZmSUN1 and ZmSUN2 are CCSD-type and are most closely related to plant SUNdomain homologs AtSUN1, AtSUN2, and OsSad1 ZmSUN3, 4, and are PM3-type and probably represent a previously ... presence and localization of ZmSUN3/4 proteins in planta, we developed peptide antibodies for western blotting and immunolocalization, and the results are summarized in Figure and The peptides used and ... LAVIFLSFVF ACLALAKLSI Figure ZmSUN4 cDNA and protein features (A) ZmSUN4 (genotype W23) full-length cDNA, showing the 5’ and 3’ UTRs, open reading frame (ORF), and poly-A tail A diagram of the protein...

Ngày tải lên: 11/08/2014, 11:21

... were tested for specificity by RTPCR and also by qRT- PCR, followed by a dissociation curve and agarose gel electrophoresis Real-time PCR conditions and analysis PCR reactions were performed in ... brizantha qRT- PCR experiments Vegetative and reproductive tissues of apomictic and sexual B brizantha were tested The relative transcription levels of the genes were determined in ovaries and anthers ... material and equivalent efficiency of cDNA synthesis from total RNA samples Based on DNA analysis by agarose gel electrophoresis and the dissociation curves (additional file 1), one single PCR product...

Ngày tải lên: 12/08/2014, 03:20

... target and novel visual stimuli in children of alcoholics Alcohol 1998; 15:119-36 92 Porjesz B And Begleiter H Human brain electrophysiology and alcoholism In: Tarter R and Thiel DV, Eds Alcohol and ... both active and passive P300 latency and age; in this same study, levels of active latency were associated with concentration ability, and passive latency with verbal proficiency and recall [72] ... 11:141-151 67 Polich J, Hoffman D P300 and handedness Psychophysiology 1988; 35:497-507 68 Hopper KD, Patel S, Cann TS, et al The relationship of age, gender, handedness, and sidedness to the size of the...

Ngày tải lên: 02/11/2012, 11:08

... to identical and similar residues, respectively (accession numbers: BC008662 and BC000578) The sequences encoding residues 1–225 (PRTFDC1) and 1–218 (HPRT) were amplified by PCR and inserted into ... Environment, Agricultural Sciences and Spatial Planning (to L.W and S.E.), the 4928 Swedish Research Council (to H.E and P.N.) and the Swedish Cancer Foundation (to H.E and P.N.) The Structural Genomics ... PRTFDC1 (A) monomer and (B) tetramer generated using symmetry-related molecules Interactions with GMP (C) GMP, phosphate and calcium ions are shown as sticks and spheres colored pink and black, respectively...

Ngày tải lên: 15/02/2014, 01:20

... rate constants are similar between NP and MP and 2248 between HP and HPO in accordance with the similar shapes and sizes between NP and MP and between HP and HPO The different Kd values are proportional ... HP and 20 mM GA The reaction was initiated with 10 lM SaDHNA at 25 °C and quenched with M HCl The reverse reaction generated both DHNP and DHMP HP, DHNP, and DHMP were oxidized to HPO, NP, and ... using both DHNP and DHMP as the substrates and the reverse reaction (the formation of DHNP and DHMP) with HP and GA For the forward reaction, three concentrations each for DHNP and DHMP were used...

Ngày tải lên: 19/02/2014, 00:20

... in green, and the cluster of hydrophobic residues Trp390, Trp393, and Trp394 is in red The catalytic site is enlarged in B and illustrates the conserved disulfide bridge between Cys239 and Cys216 ... gradient (broken line), and concentrations of LPL (j) and lipase activities (h) were measured in each fraction as described in the Materials and methods The upper, middle, and lower panels are the ... sequence containing Trp393 and Trp394 Fig Heparin binding of wild-type LPL and the WW mutant Wild-type LPL (left) and WW mutant (right) were applied to heparinSuperose FPLC and eluted by a linear...

Ngày tải lên: 21/02/2014, 15:20

... black and hog-like bestiality, and fox and wolf-like cunning and ferocity with wicked human thought and self-command, which Raphael has enshrined in that splendid harmony of scarlet silk and crimson ... Ivon and Ivaire, the Gascon Engelier, Gộrier and Gộrin, Bộrenger and Otho, Anseis and Salamon, and the old Gerard of Rousillon; and one by one drags them to where the archbishop lies dying And ... piles of cabbage and cauli-flowers, the rows of tin ware and copper saucepans, the heaps of maccaroni and pastes, of spices and drugs; the garlands of onions and red peppers and piles of apples;...

Ngày tải lên: 21/02/2014, 21:20

... spectra between free and DNA-bound RepA are shown as dashed lines RepA was 1.25 lM and 0, 0.2, 0.4, 0.6 and lM 1IR (A), 0, 0.4, 1.2, 2.4 and lM 1DR (B), and 0, 0.8, 1.8, 3.2 and lM 1DR-short (C) ... (MA7), and the oligonucleotides were subsequently annealed with their complementary strands, and puried as above UV Vis and CD spectroscopy CD spectra and melting curves of wild-type RepA and AEDANS-labeled, ... free in solution (A) and bound to 1IR (B), 1DR (C) and 1DR-short (D) Table S1 Fluorescence lifetimes and decay amplitudes for W94 and AEDANSC160, respectively, in free RepA and RepA bound to various...

Ngày tải lên: 07/03/2014, 04:20

... was pooled and used for RT -PCR analysis The cycle numbers were 21 and 28 for b-actin and IL-11, respectively a liver SSH library prepared from bacterial-challenged fish and a full-length cDNA clone ... bold and italics The ATTTA motifs in the 5¢- and 3¢-UTRs, the start and stop codons, the signal peptide predicted by SIGNALP 3.0 [23] and the potential N-glycosylation site (NQT) are in bold and ... infected fish, a full-length cDNA clone as well as a genomic DNA clone were isolated and sequenced, and the expression and modulation of this molecule was studied Results Cloning and characterization...

Ngày tải lên: 07/03/2014, 16:20

... disrupted promoter structure and exons I and II, exhibit impaired follicular development and ovulation [25] and impaired spermiogenesis in males [26] On the other hand, mice defective in Camk4 ... hemisphere and between cerebellum primordium and medulla oblongata (Fig 6, A1 and A2) Also border regions between metencephalon and pons with the third ventricle (arrows in Fig 6, A1 and A2) showed ... Stk33 -cDNA was amplified with expand long template PCR- system (Roche) using the forward 5¢-GAGCAAT GTCACAGTAGG-3¢ and reverse 5¢-TGCAGATGGAC TGTAGAC-3¢ primers The 1006-base-long Stk33 cDNA fragment...

Ngày tải lên: 16/03/2014, 23:20

... contained an incomplete GLUT ORF and lacked the start codon Using 5¢- and 3¢-RACE PCR, two overlapping full-length cDNA clones of 2.1 kb (GT -cDNA1 ) and 3.1 kb (GT -cDNA2 ) that shared 100% nucleotide ... exon was inferred by 5¢-RACE and the two alternate 3¢-ends of exon 12 were deduced by 3¢-RACE, and are delineated by the full-length cDNA clones, GT -cDNA1 and GT -cDNA The two putative polyadenylation ... showed that both GT -cDNA1 and GT -cDNA2 contain a 5¢-UTR of 203 bp and a coding region of 1599 bp (excluding the stop codon), but the 3¢-UTR of GT -cDNA1 is 345 bp while that of GTcDNA2 is 1238 bp...

Ngày tải lên: 17/03/2014, 03:20

... tachyplesin I and polyphemusin I share two highly hydrophilic and positively charged poles in the N- and C-terminal regions and in the turn Androctonin also has a highly hydrophilic and positively ... hydrophobic residues and the disulphide bridges, and a second polar face formed by charged Arg1, Arg7 and Arg9 side-chains (see Figs and 4) In general, BM-1 was more flexible than PG-1 and IB-367, but, ... plated b-sheet structure Androctonin has an unequal number of residues on each strand between the two bridges – five in the N-terminal strand and three in the C-terminal strand This causes a greater...

Ngày tải lên: 22/03/2014, 21:20

... tryptic soy agar (TSA) and in vivo and have demonstrated that their structures are chemically and antigenically distinct from the previously described O-chain polysaccharide [11] and capsule [13] Experimental ... phenol solution (4 h, 22 °C) and harvested by centrifugation (yield 25 g, wet weight) Isolation and purification of CPS and LPS The cells were washed with 2.5% saline (w/v) and extracted by the method ... Sephadex G-100 and lyophilized (yield mg) NMR spectroscopy NMR spectra were performed on Varian INOVA 500 and 600 MHz spectrometers using standard software NMR measurements were made at 25 °C and 60...

Ngày tải lên: 23/03/2014, 13:20

... triphosphates, and restriction endonucleases were purchased from New England Biolabs (Beverly, MA, USA) T4 DNA ligase was from Gibco BRL (Grand Island, NY, USA) Ni2+-PDC was purchased from Affiland (Affinity ... requirement for ATP, CoA, and NADPH for the activity of FAE1 KCS FAE1 KCS showed high activity with monounsaturated and saturated C16 and C18 and no activity with polyunsaturated C18:2 and C18:3 In addition, ... serum albumin as standard [29] Solubilization and purification of recombinant His6-FAE1 KCS Microsomal proteins were solubilized in the presence of 0.32 M NaCl and 0.5% Triton X-100 and a final protein...

Ngày tải lên: 24/03/2014, 03:21

... CA-5¢ Probe CA-5¢ -and 5¢-RACE PCR PCR 5¢ Primer extension RT -PCR (cardosin A) RT -PCR (cardosin A) RT -PCR (cardosin C) RT -PCR (cardosin C) RT -PCR (cardosin D) RT -PCR (cardosin D) RT -PCR (cardosin ... were evaluated in a 1% agarose gel, and equal amounts of RNA (1 lg) were used for cDNA synthesis (1st Strand cDNA Synthesis Kit for RT -PCR; Roche, Basel, Switzerland) Four sets of primers, specific ... proteolytic processing and ⁄ or degradation of storage proteins (A thaliana and Brassica napus APs, orizasin and fitepsin), in leaf senescence (At4g0446, BnU55032, and VuAP1), and in programmed cell...

Ngày tải lên: 30/03/2014, 09:20