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71.
Separation strategies based on size-selective precipitation of DNA fragments with polyethylene glycol (PEG) have been used for achieving desired DNA interval in automated sample preparation for next-generation sequencing. By varying PEG concentration, DNA fragments of different sizes can be precipitated onto surfaces of carboxyl-coated paramagnetic particles selectively, and therefore, the desired DNA interval can be obtained. However, one of the crucial points in this approach is to determine the critical PEG concentration for DNA fragment of a certain size. The aim of this work was to develop a convenient and reliable method for accurately determining the critical PEG concentration. In our method, at a fixed concentration of sodium chloride (NaCl), recovered DNA samples obtained with different PEG concentrations were directly quantified, and their concentrations as a function of the PEG concentration were fitted by the logistic function. The critical PEG value was easily and accurately determined from the fitted logistic function. The repeatability and stability of the critical PEG value were assessed, showing an excellent reliability of the method. Based on this method, critical PEG values of different-size DNA fragments were determined at different NaCl concentrations. The effectiveness of the method was also demonstrated by selective precipitation of DNA fragments. 相似文献
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Ji-Fu Wei Xiao-long Wei Qiu-Yu Chen Tian Huang Li-Ya Qiao Wan-Yu Wang Yu-Liang Xiong Shao-Heng He 《Biochimica et Biophysica Acta (BBA)/General Subjects》2006
A novel phospholipase A2 (PLA2) with Asn at its site 49 was purified from the snake venom of Protobothrops mucrosquamatus by using SP-Sephadex C25, Superdex 75, Heparin-Sepharose (FF) and HPLC reverse-phage C18 chromatography and designated as TM-N49. It showed a molecular mass of 13.875 kDa on MALDI-TOF. TM-N49 does not possess enzymatic, hemolytic and hemorrhagic activities. It fails to induce platelet aggregation by itself, and does not inhibit the platelet aggregation induced by ADP. However, it exhibits potent myotoxic activity causing inflammatory cell infiltration, severe myoedema, myonecrosis and myolysis in the gastrocnemius muscles of BALB/c mice. Phylogenetic analysis found that that TM-N49 combined with two phospholipase A2s from Trimeresurus stejnegeri, TsR6 and CTs-R6 cluster into one group. Structural and functional analysis indicated that these phospholipase A2s are distinct from the other subgroups (D49 PLA2, S49 PLA2 and K49 PLA2) and represent a unique subgroup of snake venom group II PLA2, named N49 PLA2 subgroup. 相似文献
75.
考察5种萃取体系(A:正己烷,B:正己烷/乙醇,C:正己烷/异丙醇,D:氯仿/甲醇,E:氯仿/乙醇)对小球藻(Chlorella phyrenoidosa)油脂的提取效果及藻渣成分的影响。实验结果表明:不同的萃取体系下,油脂得率为D(12.27%)、E(8.87%)、C(7.71%)、B(6.80%)、A(3.91%),藻渣蛋白含量为A(52.60%)、E(46.23%)、B(40.19%)、C(39.52%)、D(32.52%),藻渣碳水化合物含量为A(23.28%)、E(16.15%)、B(13.24%)、D(13.50%)、C(9.06%);藻渣色素含量为A(1.75%)、E(1.29%)、B(1.14%)、C(0.96%)、D(0.58%);藻渣灰分含量为D(3.63%)、E(2.94%)、C(2.23%)、B(2.25%)、A(1.48%)。综合考虑微藻生物柴油的生产及藻渣的可利用性,V(氯仿)/V(乙醇)=1是一种油脂萃取效果较好,藻渣营养成分损失较小的小球藻油脂萃取体系。 相似文献
76.
大型绿藻浒苔转化表达系统选择标记的筛选 总被引:1,自引:0,他引:1
主要研究了条浒苔对抗生素氯霉素和除草剂Basta的敏感性,以确定适合的阳性选择标记基因。应用不同浓度氯霉素(0、25、50、75、100、125μg/ml)和Basta(0、5、12.5、25、37.5、50μg/ml)对不同发育时期条浒苔细胞存活率影响进行了测定。实验结果表明:不同发育时期条浒苔对氯霉素和Basta的敏感性不同。其中最大浓度125μg/ml浓度的氯霉素在15d内对条浒苔孢子和小苗两个不同发育时期的细胞均难以达到全部杀死效果,相对存活率仍分别为1%和20%;而Basta对条浒苔孢子和小苗均具有很强的杀生作用,其中5μg/ml浓度的Basta在3d内可将条浒苔孢子全部杀死,12.5μg/ml浓度下约一周时间可以将浒苔小苗全部致死。本实验结果提示bar基因有可能成为浒苔基因工程较理想的选择标记基因。 相似文献
77.
缺硼与低温对黄瓜幼苗—些生理反应的影响 总被引:1,自引:0,他引:1
营养液培养试验表明,缺硼明显降低黄瓜幼苗植株的生长量,植株叶片细胞质膜K 的渗漏增加,细胞结构受损。此外,叶片中蔗糖和还原糖都明显积累。缺硼的上述反应在低温胁迫(7-8℃,FX25-28℃为对照)时表现更为明显。试验结果表明硼对于细胞膜的完整性具有重要的作用;充分供硼可以减轻低温对细胞膜的伤害。 相似文献
78.
The Tibetan Plateau is an essential area to study the potential feedback effects of soils to climate change due to the rapid rise in its air temperature in the past several decades and the large amounts of soil organic carbon (SOC) stocks, particularly in the permafrost. Yet it is one of the most under-investigated regions in soil respiration (Rs) studies. Here, Rs rates were measured at 42 sites in alpine grasslands (including alpine steppes and meadows) along a transect across the Tibetan Plateau during the peak growing season of 2006 and 2007 in order to test whether: (1) belowground biomass (BGB) is most closely related to spatial variation in Rs due to high root biomass density, and (2) soil temperature significantly influences spatial pattern of Rs owing to metabolic limitation from the low temperature in cold, high-altitude ecosystems. The average daily mean Rs of the alpine grasslands at peak growing season was 3.92 μmol CO(2) m(-2) s(-1), ranging from 0.39 to 12.88 μmol CO(2) m(-2) s(-1), with average daily mean Rs of 2.01 and 5.49 μmol CO(2) m(-2) s(-1) for steppes and meadows, respectively. By regression tree analysis, BGB, aboveground biomass (AGB), SOC, soil moisture (SM), and vegetation type were selected out of 15 variables examined, as the factors influencing large-scale variation in Rs. With a structural equation modelling approach, we found only BGB and SM had direct effects on Rs, while other factors indirectly affecting Rs through BGB or SM. Most (80%) of the variation in Rs could be attributed to the difference in BGB among sites. BGB and SM together accounted for the majority (82%) of spatial patterns of Rs. Our results only support the first hypothesis, suggesting that models incorporating BGB and SM can improve Rs estimation at regional scale. 相似文献
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