shotgun

枪壳 的意思

shotgun bullet鸟枪子弹

希望采纳~~~对照下贴韩文歌词自翻YouwillalwaysbemyYouwillalwaysbemy耨拿也ANGEL色喷拿行桑米苏几给黑怒噶目啦黑度You(you)内吗len怒有耨也ILoveyou(you)Causeyouwillalwaysbemy耨拿也ANGEL色喷拿行桑米苏几给黑怒噶目啦黑度You(you)内吗len怒有耨也ILoveyou(you)Causeyouwillalwaysbemy云谈也僧一嫩有但帮内僧也为给嫩要罗帮谷票金牌加录考要den考谷考谷考要den催诶哈len也吗len考罗心也给系屁len考要den茫茫看啊奈也忙买黑所几K目累到要den桥偶denKI累den工嫩啊剧买药den工里不目里喷内给嫩撒气你不KI老den内给他噶哇耨内给他噶哇米亲内噶太阳音den拿耨漫怕啦吧耨拿漫怕啦吧耨拿也ANGEL色喷拿行桑米苏几给黑怒噶目啦黑度You(you)内吗len怒有耨也ILoveyou(you)Causeyouwillalwaysbemy耨拿也ANGEL色喷拿行桑米苏几给黑怒噶目啦黑度You(you)内吗len怒有耨也ILoveyou(you)Causeyouwillalwaysbemy1234567气全拍给无录吗K无啦卡大滚度帮金亲谷啦嫩目里den也屁无嫩孙噶啦金哈嫩掐难以denlen铺谬‘难内衣len卡要几拿KI老K撒啦几内用混度nun噶要几内KI林加度亏掐拿金或系KI得内吗啊罗KI得耨奈抗撒啊罗AngelMyAngel拿米苏金给黑耨为诶目den海苏一搜气度几给黑AngelMyAngel拿米苏金给黑耨为诶目den海苏一搜气度几给黑耨拿也ANGEL色喷拿行桑米苏几给黑怒噶目啦黑度You(you)内吗len怒有耨也ILoveyou(you)Causeyouwillalwaysbemy耨拿也ANGEL色喷拿行桑米苏几给黑怒噶目啦黑度You(you)内吗len怒有耨也ILoveyou(you)Causeyouwillalwaysbemy无死米帮桥内衣噶也囊桥NomatterwhattheysayImmaholdyoucloser无死米帮桥内衣噶也囊桥NomatterwhattheysayImmaholdyoucloserUsedtobeasillybum气亲得录气桥den哼den罗死罗叫den内给奈给len谈啦剧Usedtobeasillybum气亲得录气桥den哼den罗死罗叫den内给奈给len谈啦剧耨拿也ANGEL色喷拿行桑米苏几给黑怒噶目啦黑度You(you)内吗len怒有耨也ILoveyou(you)Causeyouwillalwaysbemy耨拿也ANGEL色喷拿行桑米苏几给黑怒噶目啦黑度You(you)内吗len怒有耨也ILoveyou(you)Causeyouwillalwaysbemy耨拿也ANGEL色喷拿行桑米苏几给黑怒噶目啦黑度You(you)内吗len怒有耨也ILoveyou(you)Causeyouwillalwaysbemy耨拿也ANGEL色喷拿行桑米苏几给黑怒噶目啦黑度You(you)内吗len怒有耨也ILoveyou(you)Causeyouwillalwaysbemy

csol双管枪Stoger Double Defence Shotgun怎么得？属性怎么样？ 【伤害：85】【连射：17】【稳定：37】【命中：100】【后座：33】【重量：80】 对这把枪试过之后我简要发表一下自己的看法，不是强迫症千万不要去用C币买，就像奥尼玛这样的强迫症患者就毫不犹豫的用C币去解锁了，这把枪可能有的骚年觉得打生化会很不错，但是实战里面真心没有M870和AM12好用，因为换弹的速度实在太慢了，如果没有人掩护你一发打出去肯定是自身难保的，在人比较少的房间里面玩玩溜尸还行，因为伤害击退定身效果还不错，个人看法仅供参考！ 这把枪最大的亮点就是可以代替M3在躲猫猫模式里面购买使用！ PS：这把枪是直接出在了解锁系统里面，可能也就意味着他除了通过用C币直接解锁还能够用击杀条件解锁，根据解锁系统里面可以看到解锁条件是用M3达成2000击杀，经常玩躲猫猫的话迟早能解锁掉。

歌曲名:Shotgun Blues (BBC in Concert)歌手:Dr Feelgood专辑:Dr Feelgood - BBC In Concert (1st November 1978)　 I got the shotgun bluesShotgun bluesI said I don"t know what I didBut I know I gotta moveI got the shotgun bluesShotgun bluesI can"t wait here foreverI got too much here to loseAn now you"re blowin" smokeI think you"re one big jokeMe...I gotta lot to learnAn I"m still waitin" for the heads to turnYou say I walk a lineFuck they move it every timeYou walk a mile in my shoesAnd then you tell me "bout singin" the bluesYou get what you pay forAn freedom"s real high pricedAn while your rippin" off childrenSomebody"s fuckin" your wifeIt"s never made a differenceIt"s only how you surviveI got the shotgun bluesShotgun bluesI said I don"t know what I didBut I know I gotta moveI got the shotgun bluesShotgun bluesI can"t wait here foreverI got too much here to loseAn now you ask me whyI said it"s do or dieI"ll stick it right in your faceAnd then I"ll put you in your motherfuckin" placeAnd you...you can suck my assAn I think it"s so low classMe...I"m just so concernedI"m still waitin" for your ass to burnOoooh you want a confrontationI"ll give you every fuckin" chanceWith your verbal masturbationMe...I just like to danceHow"s that for provocationI"m just makin" a stanceAn I"m tired of the frustrationOf livin" inside of your liesAnd I"m wired on indignationI said somebody"s got to dieI got the shotgun bluesShotgun bluesI said I don"t know what I didBut I know I gotta moveI got the shotgun bluesShotgun bluesI can"t stay here foreverI got too much here to loseI got the shotgun bluesShotgun bluesListen motherfuckerYou"re about to pay your duesI got the shotgun bluesShotgun bluesIf you"re goin" up against meThen you know you"re gonna loseI knowI know (there"s ways- that we- can find)I knowI knowYou think anyone with an I.Q. over 15Would believe your shit...fuckheadNothin" but a fuckin" pussyhttp://music.baidu.com/song/10441850

去Minecraft中文下载站等网站下载“元素枪Mod”文件，选择对应版本。将下载的mod放置到Minecraft程序“.minecraft”——“mods”文件夹中。启动游戏，打开Mods选项查看确认mod载入成功。进入游戏，在工作台中按照枪的合成公式合成各类枪。装备枪后，配合土元素弹药就可以发射了，也可以直接使用。

pump action shotgun泵行动猎枪很高兴为您解答如果你对这个答案有什么疑问，请追问

能。ShotgunRV是一款能够提供专业强大的视频后期处理软件，可支持中文、英文切换使用。ShotgunRV软件不仅能够对视频进行多种后期操作，还能够提供行业标准图像支持，支持多格式文件播放，甚至包括立体3D。

把基因组长长的序列打断(shotgun sequencing)。短序列组装（Sequence assembly）几乎是近年来next-generation sequencing最热门的话题。简单来说，就是把基因组长长的序列打断(shotgun sequencing)。因为我们不知道基因组整条序列是如何排列（成一条链，最后成为一条染色体）组合（如何区分不同染色体）的，而我们又无法实现一次，把整条长序列完整测序（现在有单子测序可能是一个新的sunlight)。然后，我们通过算法，计算机的帮助，把这些短的序列组装起来成为一条完整有序的序列。

Get out your guns, battles begun, are you a saint, or a sinner?——持枪上膛，战役已经打响。你是一个圣人，还是一个罪人？If loves a fight, than I shall die,with my heart on a trigger——如果爱是一场战役，那我也会拼尽全力扣下扳机，即便我会因此死去。They say before you start a war, you better know what you"re fighting for.——他们说“在你开始一场战役前，你最好知道自己为何而战。”Well baby, you are all that I adore, if love is what you need, a soldier I willbe.——宝贝，你是我的挚爱，如果爱是你所要的，那我将成为你披上戎装。I"m an angel with a shotgun, fighting til" the wars won, I don"t care if heaven won"t take me back.——我是个持枪上膛的天使，为了胜利奋战到最后，我不在乎天堂是否会就此抛弃我。I"ll throw away my faith, babe, just to keep you safe.——我将抛弃我的信仰，宝贝，仅仅是为了你的安全。Don"t you know you"re everything I have?——你是否知道，你是我所拥有的一切？Sometimes to win, you"ve got to sin,don"tmean I"m not a believer.——彼时胜利，你被定罪，并不意味着我不是信徒。and I"m gonna hide, hide, hide my wings，tonight.——今晚，我将藏起我的羽翼。

歌曲名:Shotgun Blues歌手:Bob Campbell专辑:Times Ain"t Like They Used To Be, Vol. 1　 I got the shotgun bluesShotgun bluesI said I don"t know what I didBut I know I gotta moveI got the shotgun bluesShotgun bluesI can"t wait here foreverI got too much here to loseAn now you"re blowin" smokeI think you"re one big jokeMe...I gotta lot to learnAn I"m still waitin" for the heads to turnYou say I walk a lineFuck they move it every timeYou walk a mile in my shoesAnd then you tell me "bout singin" the bluesYou get what you pay forAn freedom"s real high pricedAn while your rippin" off childrenSomebody"s fuckin" your wifeIt"s never made a differenceIt"s only how you surviveI got the shotgun bluesShotgun bluesI said I don"t know what I didBut I know I gotta moveI got the shotgun bluesShotgun bluesI can"t wait here foreverI got too much here to loseAn now you ask me whyI said it"s do or dieI"ll stick it right in your faceAnd then I"ll put you in your motherfuckin" placeAnd you...you can suck my assAn I think it"s so low classMe...I"m just so concernedI"m still waitin" for your ass to burnOoooh you want a confrontationI"ll give you every fuckin" chanceWith your verbal masturbationMe...I just like to danceHow"s that for provocationI"m just makin" a stanceAn I"m tired of the frustrationOf livin" inside of your liesAnd I"m wired on indignationI said somebody"s got to dieI got the shotgun bluesShotgun bluesI said I don"t know what I didBut I know I gotta moveI got the shotgun bluesShotgun bluesI can"t stay here foreverI got too much here to loseI got the shotgun bluesShotgun bluesListen motherfuckerYou"re about to pay your duesI got the shotgun bluesShotgun bluesIf you"re goin" up against meThen you know you"re gonna loseI knowI know (there"s ways- that we- can find)I knowI knowYou think anyone with an I.Q. over 15Would believe your shit...fuckheadNothin" but a fuckin" pussyhttp://music.baidu.com/song/15146722

歌曲发行时间：2011-08-23所属专辑：《Symphony Soldier》歌手：The CabThe Cab是一支来自拉斯维加斯的流行朋克摇滚乐队，组建于2004年。2011年8月第二张专辑《Symphony Soldier（交响士兵）》问世，随即被AbsolutePunk网站评为“大作”。歌词：An angel with a shotgun, shotgun, shotgun,An angel with a shotgun, shotgun, Get out your guns, battles begun,are you a saint, or a sinner?If loves a fight, than I shall die,with my heart on a trigger.They say before you start a war,you better know what you"re fighting for.Well baby, you are all that I adore,if love is what you need, a soldier I will be.I"m an angel with a shotgun,fighting til" the wars won,I don"t care if heaven won"t take me back.I"ll throw away my faith, babe, just to keep you safe.Don"t you know you"re everything I have?..and I, wanna live, not just survive, tonight.Sometimes to win, you"ve got to sin,don"t mean I"m not a believer...and major Tom, will sing along.Yeah, they still say I"m a dreamer.They say before you start a war,you better know what you"re fighting for.Well baby, you are all that I adore,if love is what you need, a soldier I will be.I"m an angel with a shotgun,fighting til" the wars won,I don"t care if heaven won"t take me back.I"ll throw away my faith, babe, just to keep you safe.Don"t you know you"re everything I have?..and I, wanna live, not just survive, tonight.ooooohhhhhhhh, ooooooohhhhhwhoa whoa oooh whoaI"m an angel with a shotgun..fighting til" the wars won..I don"t care if heaven won"t take me back..I"m an angel with a shotgun,fighting til" the wars won,I don"t care if heaven won"t take me back.I"ll throw away my faith, babe, just to keep you safe.Don"t you know you"re everything I have?..and I, want to live, not just survive, tonight.not just ..and I"m gonna hide, hide, hide my wings tonight.They say before you start a war,you better know what you"re fighting for.Well baby, you are all that I adore,if love is what you need, a soldier I will be.

I call shotgun 我坐前排

Get out your guns, battles begun, are you a saint, or a sinner?——持枪上膛，战役已经打响。你是一个圣人，还是一个罪人？If loves a fight, than I shall die,with my heart on a trigger——如果爱是一场战役，那我也会拼尽全力扣下扳机，即便我会因此死去。They say before you start a war, you better know what you"re fighting for.——他们说“在你开始一场战役前，你最好知道自己为何而战。”Well baby, you are all that I adore, if love is what you need, a soldier I willbe.——宝贝，你是我的挚爱，如果爱是你所要的，那我将成为你披上戎装。I"m an angel with a shotgun, fighting til" the wars won, I don"t care if heaven won"t take me back.——我是个持枪上膛的天使，为了胜利奋战到最后，我不在乎天堂是否会就此抛弃我。I"ll throw away my faith, babe, just to keep you safe.——我将抛弃我的信仰，宝贝，仅仅是为了你的安全。Don"t you know you"re everything I have?——你是否知道，你是我所拥有的一切？Sometimes to win, you"ve got to sin,don"tmean I"m not a believer.——彼时胜利，你被定罪，并不意味着我不是信徒。and I"m gonna hide, hide, hide my wings，tonight.——今晚，我将藏起我的羽翼。资料：http://wenda.haosou.com/q/1398917798067366

angelbeats

shotgunrv倍速播放步骤。1、在Shotcut界面中在播放列表中导入视频素材。2、把导入的素材放在相应的轨道上。3、依次在轨道上选中该素材后鼠标右键调出菜单中选择属性。4、打开属性面板对话框中在速度栏输入速度的值。5、最后鼠标单击空白处即可。

歌曲名:Shotgun歌手:Don Byron专辑:Do the Boomerang: The Music of Junior WalkerLyrics：Ami　Music：Yoshinao Mikami　Arrangement：Dreadstore Cowboy谁だよ　袖つかんで飞び立つ俺を止めるのはやめろよ　谕すように手顷な梦を食べさせんな谁でも　怖いもんだまだ见ぬ先の手探りは言い訳　并べたって守るだけが人生じゃない痛みだって感じりゃいいさでかくなった证だろ他人ばっか眺めてないで时间に优しさはないやり直せない　my momentその1秒がひどく欲しいどれもこほせない　teacup注いだのは俺の全て笑えよ　无谋だって　壁の向こうで束になって気づけよ　胸の中　眠れる狮子が泣いてるぜ理想ばっか并べりゃいいさ梦を待ってシンデレラ波の乘って　流されないで自分の足で步け辉かせたい　my momentその1秒に震えたい热く煮えたぎる　teacup溢れそうな俺の全てここからそう远くはないだろうこのまま速度上げていいだろうじわじわ込み上げる魂よ

1、先开启这三个选项：ScreeningRoomforRV，ShotgunReview，ShotgunIntegration。2、默认拍屏上传后，视频存入Shotgun云端，这样shotgunrv就能打开mov了。

歌曲名:shotgun歌手:liberty x专辑:xRide shotgunCome onOhI don"t even think I can take no moreI know we need a place to goTime to paint the spotWe ain"t hit the floorYou know our money"sRunning real lowSo we gotta do what we gotta doYou know we gotta get that buzzUnderstand whyI don"t expect you tooNobody does it like we doLeave the City ofAngels behind usYou and me nowFeeling freeI"ll let you ride shotgunOut of L.A. todayIt"s gonna be a hot oneTime to escape and say that you wannaGo far awayDon"t want another hold up todayYou know you"re riding shotgunAll of the way to VegasWe"re gonna slot oneNo other way to say thisWe"re gonna stack up the chipsWe"re gonna be the names on your lipsYou know you"re riding shotgunEverything is so possibleIt"s like I"m living out my dreamsNow I"m feeling kinda unstoppableNothing"s ever what it seemsGotta take my chancesCos I"m here to winYou know the stakes are getting real highGo ahead and give it another spinMe I"m trying to reach the skyNobody knows what would have been(Would have been)If you never said those words to meI"ll let you ride shotgunOut of L.A. todayIt"s gonna be a hot oneTime to escape and say that you wannaGo far awayDon"t want another hold up todayYou know you"re riding shotgunAll of the way to VegasWe"re gonna slot oneNo other way to say thisWe"re gonna stack up the chipsWe"re gonna be the names on your lipsYou know you"re riding shotgunOh, ohNobody knows what would have been(Nobody knows)If you never said those words to meI"ll let you ride shotgun(Shotgun)Out of L.A. todayIt"s gonna be a hot one(Hot one)Time to escape and say that you wannaGo far away(Go far away)Don"t want another hold up todayYou know you"re riding shotgunAll of the way to VegasWe"re gonna slot oneNo other way to say this(Ah)We"re gonna stack up the chipsWe"re gonna be the names on your lips(Ooh)You know you"re riding shotgunI"ll let you ride shotgunOut of L.A. todayIt"s gonna be a hot oneTime to escape and say that you wanna(Yeah)Go far away(Go far away)Don"t want another hold up todayYou know you"re riding shotgunSlot oneStack up the chipsNames on your lipsYou know you"re riding shotgunhttp://music.baidu.com/song/14511855

筷子

有关shotgun marriage的解释如下：名词 n.1.(主美)强迫婚姻，特指男方被迫与某一被自己损害了名声的女人结婚。2.（主中国）拉郎配，强拉男子与女子结婚 奉子成婚

shotgun wedding [英]u02c8u0283u0254tu02ccgu028cn u02c8wediu014b [美]u02c8u0283ɑtu02ccɡu028cn u02c8wu025bdu026au014b n. 强制婚姻，勉强的妥协 [例句]My friend announced her shotgun wedding this morning.今天早上我的朋友宣布她要奉子成婚。

这首歌很好听

马上注册

中文名称：带枪的天使外文名称：Angel With A Shotgun所属专辑：Symphony Soldier歌曲时长：03：43发行时间：2011年8月歌曲原唱：The CabThe Cab是一支来自拉斯维加斯的流行朋克摇滚乐队，组建于2004年。2011年8月第二张专辑《Symphony Soldier（交响士兵）》问世，随即被AbsolutePunk网站评为“大作”。

目录 根据分析对象和实验目的，宏基因组的研究基本上可以分为 1. Pre-processing 2. Sequence analysis 包括两种分析策略： read-based (mapping) 和 assembly-based 简单来说，assembly-based approach 受到覆盖度的制约，因为组装时低覆盖度的区域是不会进行组装的，而是被丢弃，这样低丰度的细菌的信息就被丢弃了，反映在reads利用率上，就是往往reads利用率极低，往往低于50% 而 read-based (mapping) approach 则受到reference databases的制约，因为细菌的遗传多样性很高，即便是同一个菌种，它的不同菌株，其基因组的组成也是有相对比较大的差异的，那么在mapping的时候就会出现mapping不上的问题，使得mapping效率不够高；而且只能分析reference databases中有的物种，对于reference databases未收录的新物种，是无法进行分析的。 不过可用的微生物参考基因组正在迅速地增加，包括那些原先难以培养的细菌由于培养方法的改进，使得对其进行测序成为可能，再加上单细胞测序的途径和 metagenomic assembly的途径得到的基因组序列。现在一些类型的环境样品（如人肠道）的参考基因组的多样性已经可以满足 assembly-free taxonomic profiling 的要求。 随着测序成本的下降和测序深度的增加，其分析难度将会越来越大，制约效应也将会越来越明显 预计的单位测序成本将会以指数关系下降,但其中计算成本下降的幅度会远慢于测序成 在数据存储和数据处理的层面上，rDNA和扩增序列的分析难度较小，基本可以在个人电脑或者小型服务器上完成，但宏基因组全测序的分析却主要受限于计算技术的发展 即使在同一个环境中获取的不同样本，其微生物组成也会存在比较大的差异，这使得在样本集之间，寻找具有统计学显著性和生物学意义的差异变得很困难。因此如何做到，在即使其影响因素的作用程度很小的情况下，也能有效地检测出差异就显得十分重要。 一种策略是，构造 pilot data，即将不同浓度的绝对定量 control (spike-in) 加入到样本中，来评估实验与分析方法的稳健性(robust)； 另一种策略：two-tiered approach，即挑取少部分样本，既做 16s rDNA 测序，又做 shotgun metagenomics 测序，对比这两个层次的结果来评估实验结果的稳健性。 两种研究策略： 由于在研究宏基因组过程中，比如研究人类的微生物群，影响其微生物群的因素众多，包括宿主基因型，年龄，饮食习惯等等，当进行两个环境微生物群横向比较时，很难做到控制变量，使得在进行比较分析时混入了许多干扰因素；此时如果进行单一环境微生物群多时间点采样的纵向比较，就可以从很大程度上消除这种影响。 1. 样本量与测序深度 当实验目的是检出显著性差异时，样本量与测序深度的选择取决于（1）不同样本间微生物组组成的一致性，（2）样本固有的微生物多样性，（3）影响因素的效应量(effect size) 建议：参考前人在类似环境中的研究。若没有可参照的类似研究，选择marker gene做预实验 2. Confounding variables and control groups 在进行宏基因组研究时，往往很难找到与目标样本集对应的没有其他干扰因素的对照组 建议：目前最佳的解决策略是，尽可能地搜集各个样本群体的元数据 (metadata)，然后在随后的比较分析中将它们考虑进去。比如临床样本，包括性别、年龄、是否使用抗生素/药物、取样位置、饮食习惯等等。比如环境样本，包括地理位置、季节、pH、温度等等。 元数据的搜集可以参照MIMARKS (Minimum information about a marker gene sequence) 和 MIxS (minimum information about any (x) gene sequence) 标准 3. Sample collection/preservation 样本的处理和保存过程的差异会带来系统偏差，比如when samples are provided from a number of locations by different research groups，或者在纵向研究中，不同取样时间点的样本的保存时间长短不一。有时这些处理步骤的效应量可能比你感兴趣的生物学变量还大。 建议：尽可能按照相同的标准来进行取样和保存 4. Biomass/Contamination 当前采用的基于测序的方法具有很高的灵敏度 (highly sensitive)，即使非常微量的DNA也能被检测出来。而实验室中使用到的常规仪器和试剂并不是无菌的，这样就很可能在实验操作过程中，人为地引入污染。由于检测方法的高灵敏度，当原样本的微生物量很少时，污染带来的信号很可能会盖过真实的信号。 建议：在上机测序前，做好微生物量的定量 (qPCR)。当样品中的微生物数量少于10 5 数量级时，其极有可能会受到背景污染的干扰。此时，可以参照以下的方法进行细胞/DNA的富集： 可以增设负对照实验 (Negative control)，对其进行与实际样本相同的操作，使用相同的试剂，以此来找出污染的细菌类型，这样就可以在后续的生物信息学分析过程中将其过滤掉。 5. 选择合适的DNA提取方法 DNA提取的效果会直接对后续的实验和分析产生巨大的影响。DNA提取方法的选择依赖于样品中细胞类型的组成，然而即使是相同类型的样品其微生物组成也具有较大的差异（当人粪便中革兰氏阴性菌主导时，细胞很容易裂解，而当由相对顽强的革兰氏阳性菌主导时，则相反）。 因此不存在适用于所有样品的最佳的DNA提取方案。 若方案选择不当，则获得的DNA主要来自于那些易裂解的细菌 建议： Illumina测序仪通量大 (up to 1.5 Tb per run)，且准确率高 (with a typical error rate of 0.1–1%)，通过在不同样本的序列上添加两重barcode，可以一次测序多个samples。 然而，Illumina测序仪存在carryover (between runs) 和 carry-between (within runs)的问题。最新的测序仪由于使用了新的扩增方法 (ExAmp)，导致较高比例的‘index hopping"。 虽然没有一个明确的指导意见，告诉你在哪个特定的环境样品中应该测多大的覆盖度，但是一个基本的原则就是通量要尽可能地大，这样低丰度的细菌也能被测到。Illumina HiSeq 2500/4000， NextSeq 和 NovaSeq 的测序通量都很大，都适用于 metagenomics 的研究。 Metagenome de novo assembly 采用的策略与 whole-genome assembly 相同，均为 de Bruijn 图方法 用 de Bruijn 图方法进行宏基因组的从头组装时，面临着以下的挑战： 当进行单一基因组的组装时，其有一个前提假设：整个基因组的测序覆盖度是相对均匀的，这样就可以利用覆盖度信息来识别重复序列和鉴定测序错误和等位变异。 而metagenome中，各个组成基因组的覆盖度取决于它们的物种丰度，低丰度物种的基因组就会由于总体测序深度不够而使得最终组装出来的基因组是支离破碎的。使用更短的 k-mer 有助于低丰度基因组的组装，但是这会使得图中重复 k-mer 的频率大大增加，降低了组装的准确性。 这需要组装工具在考量低丰度物种与获得高丰度物种更长更准确的contig之间进行权衡，即选择合适的 k-mer ： 同种细菌的不同菌株，它们的基因组组成很相近，常常就是一个碱基的变异或者整个基因/操纵子的丢失，当进行 de Bruijn 图组装时，就会在这些差异的位置出现分叉，组装工具在遇到这些分叉时，常常会停在这些位置，从而导致一个个不连续组装片段的产生。 Meta-IDBA：将图依据其拓扑结构拆分成各个元件，每个元件代表各个亚种的共有区域 解决计算能力与内存不足的策略： Metagenome 组装完成后，我们得到的是成千上万的 contigs，我们需要知道哪些 contigs 来自哪一个基因组，或者都有哪些微生物的基因组。所以需要将 contigs 按照物种水平进行分组归类，称为 "bining" 一个很容易想到的策略就是，将组装得到的片段与已知物种的参考基因组进行比对，根据同源性进行归类。然而目前大多数的微生物的基因组还没有测序出来，因此限制了这种方法的可行性。 目前主流的 bining 策略利用的是 contigs 的序列组成特点。 依据：来自同一菌株的序列，其核酸组成是相似的 例如根 据核酸使用频率 （oligonucleotide frequency variations），通常是四核苷酸频率（tetranucleotide frequency）， GC含量 和 必需的单拷贝基因 等 优势：即便只有一个样品的宏基因组数据也可以进行binning，这在原理上是可操作的 不足：由于很多微生物种内各基因型之间的基因组相似性很高，想利用1个样品的宏基因组数据通过核酸组成信息进行binning，效果往往并不理想或难度很大。利用核酸组成信息进行binning，基本上只适合那些群落中物种基因型有明显核酸组成差异的，例如低GC含量和一致的寡核苷酸使用频率 依据：来自同一个菌株的基因在不同的样品中 ( 不同时间或不同病理程度 ) 的丰度分布模式是相似的【PMID: 24997787】。 原因：比如，某一细菌中有两个基因，A和B，它们在该细菌基因组中的拷贝数比例为 A:B = 2:1，则不管在哪个样品中这种细菌的数量有多少，这两个基因的丰度比例总是为 2:1 优势：这种方法更有普适性，一般效果也比较好，能达到菌株的水平 不足：必须要大样本量，一般至少要50个样本以上，至少要有2个组能呈现丰度变化 ( 即不同的处理，不同的时间，疾病和健康，或者不同的采样地点等 ) ，每个组内的生物学重复也要尽量的多 对于像质粒这样的可移动遗传单元 (mobile genetic elements (MGEs))，由于其复制独立于细菌染色体，则同一种细菌的不同个体，该质粒的拷贝数可能存在差异，使得无法用丰度信息进行有效地bining 将核酸组成信息和丰度差异信息创建一个综合的距离矩阵，既能保证binning效果，也能相对节约计算资源，现在比较主流的binning软件多是同时依据核酸组成和丰度变化信息 依据：不同的细菌，其基因组甲基化模式不同，平均一种细菌有3种特意的甲基化 motif。MGEs (mobile genetic elements) 中含有 MTase 基因，其基因水平转移是细菌甲基化组多样性的驱动因素。虽然 MGEs 在不同个体的拷贝数不同，但是都存在，因此具有相同 MGEs 的细菌个体，其总遗传物质（包括染色体和 MGEs ）都会受到相同的MTase的作用而得到相同的甲基化模式。 Q1：从哪些序列下手进行binning呢？ 从原始的clean reads，还是从组装成的contig，还是从预测到的gene，都可以。根据基于聚类的序列类型的不同，暂且分为reads binning， contig binning和 genes binning 比较这三种binning的优劣： 总体来说应用最广泛的就是基于genes binning 和 contig binning Genes binning的一般流程 在宏基因组做完组装和基因预测之后，把所有样品中预测到的基因混合在一起，去冗余得到unique genes集合，对这个unique genes集合进行binning，主要是根据gene在各个样品中的丰度变化模式，计算gene之间的相关性，利用这种相关性进行聚类 该图中的聚类过程类似于 K-means聚类 ：随机选择几个seed genes作为诱饵，计算其他基因丰度分布模式与seed genes的相关性，按照固定的相关性值PCC>0.9，将它们归属于不同seed genes所代表的类，然后在聚好的类内重新选择seed genes，进行迭代，最终聚类得到一个个基因集合，较大的集合（超过700个基因）称为 metagenomic species (MGS)，较小的集合称为 co-abundance gene group (CAG) 基于 bining 结果进行单菌组装： 比如对核酸组成信息的利用，开发得就不够充分，四碱基使用频率因简单而被广泛使用和接受，但现在已有研究表明k-mer丰度信息也是很好的种系特征，同时越长的k-mer含有越多的信息，还有基因和参考基因组间的同源关系也是有价值的种系信号，但这些都还没有被自动化的binning软件整合 想要获得高质量的bins经常需要手动调整 Taxonomic profiling: identifies which microbial species are present in a metagenome and estimates their abundance 优点： 当然它也有局限性： 对于与人类密切相关的样品，比如人肠道，可以使用该策略，而且已经有相关的成功实践 By looking at co-abundant markers from preassembled environment-specific gene catalogs 即前人研究 (MetaHIT consortium) 已经得出特定环境下的微生物的组成，这些微生物中有某些 co-abundant markers（这些 marker genes 的丰度与其物种的丰度成正比），这样就可以基于对这些 markers 的定量得到对应的物种丰度 选择 markers 的不同策略： 当样本量巨大，都进行组装是明显不切实际的，此时采用 marker-based approaches 是一个不错的选择；而且，如果该环境来源的样本其组成微生物是研究比较充分时，marker-based approaches 能得到比较准确的物种定量结果。 Gene identification Characterization of the functional potential of the microbiome 局限性 ： lack of annotations for accessory genes in most microbial species 因为在评估微生物群体的代谢潜能时，只对那些高度保守和 housekeeping 类型的功能进行了注释，这就解释了，为什么来自不同环境的不同样品，它们的功能特征常常是十分相似的，即使它们的物种组成有很大差异。 例如，鉴定出微生物群落中的抗生素抗性基因，该方法高度依赖特定功能相关基因集注释的质量。 参考资料： (1) 魏子艳, 金德才, 邓晔. 环境微生物宏基因组学研究中的生物信息学方法[J]. 微生物学通报, 2015, 42(5):890-901. (2) Quince C, Walker A W, Simpson J T, et al. Shotgun metagenomics, from sampling to analysis[J]. Nature Biotechnology, 2017, 35(9):833. (3) 句句干货！一文读懂宏基因组binning (4) Nielsen H B, Almeida M, Juncker A S, et al. Identification and assembly of genomes and genetic elements in complex metagenomic samples without using reference genomes[J]. Nature Biotechnology, 2014, 32(8):822-828. (5) Sangwan N, Xia F, Gilbert J A. Recovering complete and draft population genomes from metagenome datasets[J]. Microbiome, 2016, 4(1):8. (6) Abubucker, S. et al. Metabolic reconstruction for metagenomic data and its application to the human microbiome. PLoS Comput. Biol. 8, e1002358(2012). (7) Beaulaurier J, Zhu S, Deikus G, et al. Metagenomic binning and association of plasmids with bacterial host genomes using DNA methylation.[J]. Nature Biotechnology, 2017, 36(1).

歌曲名:Shotgun歌手:Richard Elliot专辑:CrushThe Rasmus - ShotProducer:AQTonight we escapeJust you and meWe"ll find our peaceSomewhere across the seasEnough of the frightEnough of the fussI"ll be awake if he finds usNeedless to sayI"ll stand in your wayI will protect youAnd I...I"ll take the shot for youI"ll be the shield for youNeedless to sayI"ll stand in your wayI"ll take the shot for youI"ll give my life for youI"ll make it stopI"ll take the shotFor youFor youTonight we"ll be freeI"ll find us a homeTonight we will beFinally on our ownEnough of the hellEnough of the painI won"t let him touch youI love youNeedless to sayI"ll stand in your wayI will defend youAnd I...I"ll take the shot for youI"ll be the shield for youNeedless to sayI"ll stand in your wayI"ll take the shot for youI"ll give my life for youI"ll make it stopI"ll take the shotFor youFor youEnough of the scarsEnough broken heartsI will protect youAnd I...I"ll take the shot for youI"ll be the shield for youNeedless to sayI"ll stand in your wayI"ll take the shot for youI"ll give my life for youI"ll make it stopI"ll take the shotI"ll take the shotI"ll make it stopI"ll make it stopI"ll take the shothttp://music.baidu.com/song/8263760

歌曲名:Shotgun歌手:Tony Lucca专辑:ShotgunSweetbox - Shot GunBOOM SHOT GUN AMMUNITION ITS COMING IN MY DIRECTIONI"M THE TARGET ON THE RUN GOT NO DAMAGE TO BE DONEI"M THE NUMBER IN YOUR COLLECTIONBET YOU WOULD LOVE ME TO STAYBET YOU WOULD LOVE ME TO PLAYSHOT GUN BABY YOUR A SHOT GUNLOOKING FOR SELECTION TRYNA SHOOT ME DOWN DOWN DOWNSHOT GUN RUNNING ON A MISSIONCRITICAL CONDITION TRYNA SHOOT ME DOWN ME DOWN ME DOWNDONT MESS WITH ME (DONT MESS WITH ME)DONT MESS WITH ME (DONT MESS WITH ME)BOOM SHOT GUN NOT MY TYPE OF FUN YOU CAN REST YOUR POSITIONI"M NOT YOUR MIDNIGHT GAME I DONT RELOCATE YOUR AIMI"M OUT OF YOUR COMPETITIONBET YOU WOULD LOVE ME TO STAYBET YOU WOULD LOVE ME TO PLAYSHOT GUN BABY YOUR A SHOT GUNLOOKING FOR SELECTION TRYNA SHOOT ME DOWN DOWN DOWNSHOT GUN RUNNING ON A MISSIONCRITICAL CONDITION TRYNA SHOOT ME DOWN ME DOWN ME DOWNDONT MESS WITH ME (DONT MESS WITH ME)DONT MESS WITH ME (DONT MESS WITH ME)GO AHEAD TAKE YOUR SHOT TO MEGIVE IT OFF BET YOU"VE GOT YOU"LL MISS MESHOT GUN BABY YOUR A SHOT GUNLOOKING FOR SELECTION TRYNA SHOOT ME DOWN DOWN DOWNSHOT GUN RUNNING ON A MISSIONCRITICAL CONDITION TRYNA SHOOT ME DOWN ME DOWN ME DOWNSHOT GUN BABY YOUR A SHOT GUNLOOKING FOR SELECTION TRYNA SHOOT ME DOWN DOWN DOWNSHOT GUN RUNNING ON A MISSIONCRITICAL CONDITION TRYNA SHOOT ME DOWN ME DOWN ME DOWNhttp://music.baidu.com/song/2549489

就是副驾驶位的意思啦

天使的房间你躺着，默默地在我面前你的眼泪，对我也没有意义风，在窗外咆哮你从不给予我的爱，我给你真的不该得到它 但是现在，你什么也不能做就那样睡在你惟一的记忆中而且哭泣，我最亲爱的母亲这是一首摇篮曲 闭上你的眼睛 再见这是你所唱的 虽然我从未留意我并没有觉得应该为你哭泣这是一首摇篮曲 闭上你的眼睛 再见 再见……虽然是如此细微的事情却深深地埋葬在我心里你会消失吗？ 你会离开吗？消失在地底下也许会飞到高高的云中也许没有我你会很快乐这么多的种子撒在田野上又有哪些能受祝福而发芽？假如我死了 我就不再有那样多的忧愁你将听不到我说：对不起希望之光在哪里？我怀疑它正在某处哭泣这是一首摇篮曲 闭上你的眼睛 再见这是你所唱的 虽然我从未留意我并没有觉得应该为你哭泣这是一首摇篮曲 闭上你的眼睛 再见 再见……

上纵梁。shotgun在汽车结构中通常指上纵梁，前纵梁加强结构，即铰链柱到大灯横梁之间的结构。整车前端结构中，防撞梁、吸能盒、纵梁、shotgun是主要传力路径对应的结构承载部件。

n.(名词)猎枪鸟枪<美>【橄】拉开进攻阵式霰弹枪散弹枪滑膛枪火枪散弹猎枪不得不举行的婚礼（橄榄球）进攻阵势adj.(形容词)狭长的盒子式的强迫的强制的用武力的漫无目标的任意的用猎枪的漫无目的的v.(动词)射击强迫用猎枪射击int.(感叹词)感叹词由某人喊出要求得到坐在汽车旅客席的特殊待遇

副驾驶座位,美剧中提及过

那是突然间到来数以百计的Love song在我体内呼喊著想就这样交给冲动来吧宝贝今晚也有不知从何而来 鸣响著的声音闇夜中交织著 爱的交响曲让我们也来做看看吧！想藉由手的触碰来感受著这一刻最佳的瞬间就在这 你那把炽热的散弹枪绝对不要离开喔在这里的话 你是我甜蜜的所有物(宝物}那般炽热的夜晚 也一晚便冷却下来如同魔法解除般 一瞬回复原状别那样碰触我已经没有什麼可说的了吧你也已经没有兴趣了若是开始得早的话结果也会随著更早到来吧呐最重要的时刻呀即刻消失 就如同散弹枪般呢而最小损害的是两人再次成为毫无联系的陌生人哪---music---如同破抹布般丢弃还是 被丢弃的世界呢若是彼此深爱对方的话无论谁都会受到伤害呀既然如此不如就这麼一次感情性的在战场中让它以这种方式结束好呢时有时无的自我放弃感伤性的发炎症状为了消失今天也 想感觉到来吧 正是现在真正的Love song那种东西 并不存在於这里唷在最差劲的存在中腐烂了 而我也将那样 哎哎哎哎最佳的瞬间就在这 你那把炽热的散弹枪绝对不要离开喔在这里的话 你是我甜蜜的所有物(宝物）但是若到了明天 再会了

歌曲名:Shotgun歌手:Jr. Walker & The All Stars专辑:The Motown Story: The SixtiesThe Rasmus - ShotProducer:AQTonight we escapeJust you and meWe"ll find our peaceSomewhere across the seasEnough of the frightEnough of the fussI"ll be awake if he finds usNeedless to sayI"ll stand in your wayI will protect youAnd I...I"ll take the shot for youI"ll be the shield for youNeedless to sayI"ll stand in your wayI"ll take the shot for youI"ll give my life for youI"ll make it stopI"ll take the shotFor youFor youTonight we"ll be freeI"ll find us a homeTonight we will beFinally on our ownEnough of the hellEnough of the painI won"t let him touch youI love youNeedless to sayI"ll stand in your wayI will defend youAnd I...I"ll take the shot for youI"ll be the shield for youNeedless to sayI"ll stand in your wayI"ll take the shot for youI"ll give my life for youI"ll make it stopI"ll take the shotFor youFor youEnough of the scarsEnough broken heartsI will protect youAnd I...I"ll take the shot for youI"ll be the shield for youNeedless to sayI"ll stand in your wayI"ll take the shot for youI"ll give my life for youI"ll make it stopI"ll take the shotI"ll take the shotI"ll make it stopI"ll make it stopI"ll take the shothttp://music.baidu.com/song/10281687

shotgunusetheduo的曲风？是一种电子音乐风格。

佛罗里达Sunrise市Shotgun路757号,邮编：33326 Sunrise是美国佛罗里达州的一个城市,Shotgun是路名

歌曲名:Shotgun Blues歌手:Isobel Campbell&mark lanegan专辑:Sunday At Devil Dirt　 I got the shotgun bluesShotgun bluesI said I don"t know what I didBut I know I gotta moveI got the shotgun bluesShotgun bluesI can"t wait here foreverI got too much here to loseAn now you"re blowin" smokeI think you"re one big jokeMe...I gotta lot to learnAn I"m still waitin" for the heads to turnYou say I walk a lineFuck they move it every timeYou walk a mile in my shoesAnd then you tell me "bout singin" the bluesYou get what you pay forAn freedom"s real high pricedAn while your rippin" off childrenSomebody"s fuckin" your wifeIt"s never made a differenceIt"s only how you surviveI got the shotgun bluesShotgun bluesI said I don"t know what I didBut I know I gotta moveI got the shotgun bluesShotgun bluesI can"t wait here foreverI got too much here to loseAn now you ask me whyI said it"s do or dieI"ll stick it right in your faceAnd then I"ll put you in your motherfuckin" placeAnd you...you can suck my assAn I think it"s so low classMe...I"m just so concernedI"m still waitin" for your ass to burnOoooh you want a confrontationI"ll give you every fuckin" chanceWith your verbal masturbationMe...I just like to danceHow"s that for provocationI"m just makin" a stanceAn I"m tired of the frustrationOf livin" inside of your liesAnd I"m wired on indignationI said somebody"s got to dieI got the shotgun bluesShotgun bluesI said I don"t know what I didBut I know I gotta moveI got the shotgun bluesShotgun bluesI can"t stay here foreverI got too much here to loseI got the shotgun bluesShotgun bluesListen motherfuckerYou"re about to pay your duesI got the shotgun bluesShotgun bluesIf you"re goin" up against meThen you know you"re gonna loseI knowI know (there"s ways- that we- can find)I knowI knowYou think anyone with an I.Q. over 15Would believe your shit...fuckheadNothin" but a fuckin" pussyhttp://music.baidu.com/song/10304983

歌曲名:Shotgun Blues歌手:Junior Wells专辑:Best Of The Vanguard Years　 I got the shotgun bluesShotgun bluesI said I don"t know what I didBut I know I gotta moveI got the shotgun bluesShotgun bluesI can"t wait here foreverI got too much here to loseAn now you"re blowin" smokeI think you"re one big jokeMe...I gotta lot to learnAn I"m still waitin" for the heads to turnYou say I walk a lineFuck they move it every timeYou walk a mile in my shoesAnd then you tell me "bout singin" the bluesYou get what you pay forAn freedom"s real high pricedAn while your rippin" off childrenSomebody"s fuckin" your wifeIt"s never made a differenceIt"s only how you surviveI got the shotgun bluesShotgun bluesI said I don"t know what I didBut I know I gotta moveI got the shotgun bluesShotgun bluesI can"t wait here foreverI got too much here to loseAn now you ask me whyI said it"s do or dieI"ll stick it right in your faceAnd then I"ll put you in your motherfuckin" placeAnd you...you can suck my assAn I think it"s so low classMe...I"m just so concernedI"m still waitin" for your ass to burnOoooh you want a confrontationI"ll give you every fuckin" chanceWith your verbal masturbationMe...I just like to danceHow"s that for provocationI"m just makin" a stanceAn I"m tired of the frustrationOf livin" inside of your liesAnd I"m wired on indignationI said somebody"s got to dieI got the shotgun bluesShotgun bluesI said I don"t know what I didBut I know I gotta moveI got the shotgun bluesShotgun bluesI can"t stay here foreverI got too much here to loseI got the shotgun bluesShotgun bluesListen motherfuckerYou"re about to pay your duesI got the shotgun bluesShotgun bluesIf you"re goin" up against meThen you know you"re gonna loseI knowI know (there"s ways- that we- can find)I knowI knowYou think anyone with an I.Q. over 15Would believe your shit...fuckheadNothin" but a fuckin" pussyhttp://music.baidu.com/song/2745251

歌曲名:Shotgun Blues歌手:Guns N" Roses专辑:Use Your Illusion Ii　 I got the shotgun bluesShotgun bluesI said I don"t know what I didBut I know I gotta moveI got the shotgun bluesShotgun bluesI can"t wait here foreverI got too much here to loseAn now you"re blowin" smokeI think you"re one big jokeMe...I gotta lot to learnAn I"m still waitin" for the heads to turnYou say I walk a lineFuck they move it every timeYou walk a mile in my shoesAnd then you tell me "bout singin" the bluesYou get what you pay forAn freedom"s real high pricedAn while your rippin" off childrenSomebody"s fuckin" your wifeIt"s never made a differenceIt"s only how you surviveI got the shotgun bluesShotgun bluesI said I don"t know what I didBut I know I gotta moveI got the shotgun bluesShotgun bluesI can"t wait here foreverI got too much here to loseAn now you ask me whyI said it"s do or dieI"ll stick it right in your faceAnd then I"ll put you in your motherfuckin" placeAnd you...you can suck my assAn I think it"s so low classMe...I"m just so concernedI"m still waitin" for your ass to burnOoooh you want a confrontationI"ll give you every fuckin" chanceWith your verbal masturbationMe...I just like to danceHow"s that for provocationI"m just makin" a stanceAn I"m tired of the frustrationOf livin" inside of your liesAnd I"m wired on indignationI said somebody"s got to dieI got the shotgun bluesShotgun bluesI said I don"t know what I didBut I know I gotta moveI got the shotgun bluesShotgun bluesI can"t stay here foreverI got too much here to loseI got the shotgun bluesShotgun bluesListen motherfuckerYou"re about to pay your duesI got the shotgun bluesShotgun bluesIf you"re goin" up against meThen you know you"re gonna loseI knowI know (there"s ways- that we- can find)I knowI knowYou think anyone with an I.Q. over 15Would believe your shit...fuckheadNothin" but a fuckin" pussyhttp://music.baidu.com/song/7926554

歌曲名:Shotgun Symphony歌手:Blind Stare专辑:Symphony of DelusionsShotgun SymphonyBlind StareSymphony of DelusionsBlind Stare--Shotgun Symphony--lndjove--......It all begins from this pointand it"s the point for all this to endEvery story has got an ending,but his is the oneThat keeps going without it......I"ve been haunted by the impossibleAnd blinded by my own eyesThis is the end of me!I pull the trigger to escape......I have followed the path and walked through the doorI have hated you so much, but now I knowIt would be pointless forevermoreCountless nights spent onjust pacing under the streetlightsThis is the part of the playwhere the hero is sentenced to death......This is called my realityand it is just my dreamThe hate so pure is the life of meand it is what I amEvery step towards you takes metowards to my endThe truth is now revealedand I am standing at the edge......I"ve been haunted by the impossibleAnd blinded by my own eyesThis is the end of me!I pull the trigger to escapeWaking up every morning is like entering a dreamSadness is just the gate to my own realmWalking forward is no longer an optionI am no longer part of this world......This is called my realityand it is just my dreamThe hate so pure is the life of meand it is what I amEvery step towards you takes metowards to my endThe truth is now revealedand I am standing at the edgeThis is called my realityand it is just my dreamThe hate so pure is the life of meand it is what I amEvery step towards you takes metowards to my endThe truth is now revealedand I am standing at the edgehttp://music.baidu.com/song/24782611