Calvaria was obtained and scissored into pieces on the super clean blench through sterile method after fetal Kunming mouse within 24 hours were killed by cervical dislocation in the experiment. The bone pieces were digested by 0.25% trypsin for 30 min at 37℃ at first, then digested by 0.2% collagenase at 37℃ in constant temperature oscillator for 60 min, and rinsed repeatedly to disaggregate more single-cells.

試驗采用24h內(nèi)昆明乳鼠色欲无码综合视频一区二区，拉頸處死2020人人操人人摸，無菌取出頭蓋骨瑟瑟欧，在超凈工作臺內(nèi)將其剪碎黑屌性爱视频，先用0.25%胰蛋白酶37℃消化30min成都毛片，然后用0.2%膠原酶37℃振蕩消化60min国产精品亚洲獵奇在线观看，反復吹打骨片分離更多細胞。

To investigate the mechanisms of biohydrogen production by anerobic fermentation of solid organic waste, gas chromatography is used to analysis the biogas and volatile fatty acid in anaerobic bio-reactor. Firstly, the ability of hydrogen production by digested sludge from the West Lake, Enteobacter aerogences, digested sludge from sewage farm, the fluid of methane pool and dejecta have been studied. The rule of the ability of hydrogen production by different bacteria under different control condition. Simultaneity, the importance of the synergistic effect on hydrogen production has been proved.

本文首先選用西湖底的厭氧活性污泥黄页三级、產(chǎn)氣腸桿菌人妖色偷偷福利网站、污水處理廠的淤泥、沼氣池發(fā)酵液以及豬糞等不同菌群對廢棄食物——馬鈴薯進行厭氧發(fā)酵產(chǎn)氫特性實驗研究激情小说图片亚洲，得到馬鈴薯在各不同菌群及工況下的發(fā)酵產(chǎn)氫能力一本到高清久久婷婷国产，同時發(fā)現(xiàn)產(chǎn)氫菌間的協(xié)同作用很重要，在控制好發(fā)酵條件的情況下扒开女高中粉嫩小泬免费视频，產(chǎn)氫菌群發(fā)酵通常會好于單一產(chǎn)氫菌的發(fā)酵看黄9999。

Articular chondrocytes from the knee joints of the A group were separated respectively to fragments, and then digested by enzymes, both the chondrocytes and the uncomplite digested tiny chondral tissues have been cultured in vitro for about 2 weeks.

分別取A組9只兔子的雙側膝關節(jié)股骨滑車及部分髁部的軟骨，在體外進行分離和消化日韩高清黄免费一区，未完全消化的軟骨組織加軟骨細胞在培養(yǎng)瓶內(nèi)一同培養(yǎng)2周后配制成濃度為1.6×10~8/ml的軟骨細胞懸液黄色视频在线播放国产，等分裝于兩個EP管中。

Then pMD-P80 is digested by the enzymes Xho I and Apa I, separated and linked to the linear plasmid pEGPF-C1 after digested by Xho I and Apa I. So the recombined expression plasmid pGFP-P80 is achieved. Then it is sequenced, PCR and digested by the restriction enzymes Xho I and Apa I. The results give that pGFP-P80 is achieved successfully and the insert sites, direction and reading frame are all right. It builds the basis for expressing, purifying, gaining p80 protein from mammiferous cells.

將pMD-P80 分別經(jīng)Xho I 和Apa I 雙酶切和回收国产片一区二区在线视频，然后與經(jīng)過Xho I 和Apa I 酶解的真核表達載體pEGPF-C1 連接人人干人人操超碰chinese、轉化，獲得重組質(zhì)粒二级片网站，經(jīng)PCR一級免费视频，Xho I 和Apa I 限制性酶切和序列測定，鑒定為真核表達質(zhì)粒pEGFP-P80国产一区二区男女激情视频，并且目的基因的插入位置久久久久久精品毛片A级、方向和讀碼框完全正確，為在哺乳動物細胞中表達并分離純化p80 蛋白奠定了基礎WWW免费簧片AV网站COM。

The Charon library DNA was then digested with restriction enzymes BamH Ⅰ and Hind Ⅲ, and DNA fragments of length of about 0.5-1. 5kb was subcloned into plasmid pUC 18 which was also digested by BamH Ⅰ and Hind Ⅲ and dephosphorylated by alkaline phosphatese before being ligated with Charon fragments.

抽提λCharon文庫總DNA亚洲久色，用BamHⅠ和HindⅢ完全酶切電泳，用低熔點瓊脂糖回收長約0.5-1.5kb的DNA片段鲁一鲁中文字幕www；連接到pUC18上在线看手机毛片，pUC DNA先用BamHⅠ和HindⅢ酶切，后用堿性磷酸酯酶脫磷影视先锋av在线资源。

Torenia fournieri genomic DNA was digested with DraI亚洲一区有码在线、EcoRV、PvuII、SmaI respectively. A special adaptor was ligated to the ends of the digested DNA fragments as a template for adaptor PCR. With the adaptor and TfPLC1 gene-specific primers, bands of 798bp and 813bp upstream of TfPLC1 were obtained successively.

應用銜接頭PCR技術日韩欧美综合在线，以藍豬耳全基因組DNA分別經(jīng)DraI国产乱女伦在线视频、EcoRV、PvuII人成视频在线观看免费、SmaI消化后與銜接頭連接產(chǎn)物為模板，用銜接頭引物和TfPLC1基因的特異引物經(jīng)過多輪的巢式PCR国产免费性交片，先后克隆到兩個大小為798bp日韩福利短片在线看免费视频播放网站、813bp的TfPLC1基因上游序列；經(jīng)測序国产一级A片妓女视频AⅠ换脸、blastn比較分析和拼接得到一個藍豬耳TfPLC1基因的啟動子序列国产又大又粗又硬又湿，共1432bp。

We biopsied 1-2 single blastomere from 6-8 cell cleavage stage intracytoplasmic sperm injection embryo, and using nested PCR amplified the high frequently mutation region G380R of FGFR3 gene of the single blastomere. The products of PCR were digested by restriction enzyme Bfm Ⅰ, then the digested products were detected by 10% PAGE to see whether the embryo inherted the mutation of the patient and to screen out normal embryo transfer.

活檢經(jīng)胞質(zhì)內(nèi)單精子注射(intracytoplasmic sperm injection国产一级黄色视频无码，ICSI)授精的胚胎發(fā)育至6～8細胞期的1～2個單卵裂球中文字幕三级视频大全，采用巢式PCR擴增單卵裂球的FGFR3基因的高發(fā)突變位點G380R區(qū)域，用限制性內(nèi)切酶BfmⅠ消化巢式PCR的內(nèi)擴增產(chǎn)物亚洲三级片在线网站，再經(jīng)10%聚丙烯酰胺凝膠電泳檢測有無遺傳患者的該種突變亚洲第一中文字幕，從而篩選出正常胚胎移植。

By use of site mutation strategy and PCR technology, we obtained the gene P12X3C that includes full length P1, 2A, 3C and a part of 2B and 3B and the gene P12X3C3D that includes full length P1, 2A, 3C, 3D and a part of 2B and 3B. After being digested by restriction enzyme respectively, the gene P12X3C and the gene P12X3C3D were cloned into the pcDNA3. 1 and pTARGET expression vector that were digested by the same enzyme. Recombinant plasmids were checked by restriction enzyme analysis and nucleic acid sequencing. Further more, recombinant plasmids were transfected into BHK-21 cells by using lipoid. The proteins of foot-and-mouth disease virus , which were expressed in BHK-21 cells, were confirmed by sandwich-ELISA and fluoroscopy, and the capsid of FMDV was tested by electron microscope. In order to evaluate enhanced immune response of guinea pigs against FMDV, DNA vaccines which were designed to produce viral capsids lacking infectious viral nucleic acid and contained the gene P12X3C and the gene P12X3C3D were injected respectively with FMDV 3D protein which was expressed in Pichia Pastoris Secreted expression System and purified or with pcDNA3. 1/IFN which includes the gene IFN-α of cattle. Subsequently, Recombinant plasmids were injected to cattles with or without pcDNA3. 1/IFN. Anti-FMDV antibodies were detected by ELISA, and the T lymphocyte proliferation response was tested by MTT assay, neutralization antibodies titers were analyzed by micro-neutralization assay.

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By use of site mutation strategy and PCR technology, we obtained the gene P12X3C that includes full length PI, 2A, 3C and a part of 2B and 3B and the gene P12X3C3D that includes full length PI, 2A, 3C, 3D and a part of 2B and 3B. After being digested by restriction enzyme respectively, the gene P12X3C and the gene P12X3C3D were cloned into the pcDNA3.1 and pTARGET expression vector that were digested by the same enzyme. Recombinant plasmids were checked by restriction enzyme analysis and nucleic acid sequencing. Further more, recombinant plasmids were transfected into BHK-21 cells by using lipoid. The proteins of foot-and-mouth disease virus, which were expressed in BHK-21 cells, were confirmed by sandwich-ELlSA and fluoroscopy, and the capsid of FMDV was tested by electron microscope. In order to evaluate enhanced immune response of guinea pigs against FMDV, DNA vaccines which were designed to produce viral capsids lacking infectious viral nucleic acid and contained the gene P12X3C and the gene P 12X3C3D were injected respectively with FMDV 3D protein which was expressed in Pichia Pastoris Secreted expression System and purified or with pcDNA3.1/lFN which includes the gene IFN-a of cattle. Subsequently, Recombinant plasmids were injected to catties with or without pcDNA3.1/IFN. Anti-FMDV antibodies were detected by ELISA, and the T lymphocyte proliferation response was tested by MTT assay, neutralization antibodies liters were analyzed by micro-neutralization assay.

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In experiment 2, pullets previously maintained on a folate-devoid diet for 4 weeks, were used to compare the bioavailability of PGA with natural tetrahydrofolate from digested bacterial cell powder after a single forced feeding for 4 weeks repletion.

試驗二：利用新母雞四周的葉酸耗竭與四周的補滿試驗，比較合成的氧化型喋醯麩胺酸（pteroylglutamic acid欧美乱图； PGA）與經(jīng)消化的細菌細胞粉末(digested bacterial cell powder日韩三级电影网； DBCP，含有天然的還原型四氫葉酸)日韩美国一级欧美一级，在新母雞體內(nèi)的吸收與代謝欧美高清黄色。