近期，一档父子亲情互动节目《爸爸去哪儿》热播。在电视屏幕中，郭涛对自己的的儿子采取了“纯爷们式”的教育，让“当下孩子的挫

李晓杰-朋友的酒
昨日一去不复回哦也
开心比什么都贵
覆水不能再收回哦也
桃花谢了有玫瑰
人生几十年总会有风雨来陪
潇潇洒洒赴会今不醉不归
往事后不后悔慢慢去体会
此刻朋友这杯酒最珍贵
快把酒满上干了这杯大声歌唱
好朋友好朋友今宵多欢畅
理想改变了我们的模样
也让我懂得了要珍惜朋友的肩膀
阳光总会在风雨之后撒向苍茫
让我们抬起头学会了坚强
就像朋友的这杯烈酒烫我心上
温暖我凡事别再放心上
今天我们来相会哦也
深情斟满了酒杯
岁月匆匆如流水哦也
管他昨天谁是谁
人生的旅程有苦有甜也有泪
好汉不提当年酒再干一杯
往事决不后悔不必再理会
此刻朋友这杯酒最珍贵
快把酒满上干了这杯大声歌唱
好朋友好朋友今宵多欢畅
理想改变了我们的模样
也让我懂得了要珍惜朋友的肩膀
阳光总会在风雨之后撒向苍茫
让我们抬起头学会了坚强
就像朋友的这杯烈酒烫我心上
温暖我凡事别再放心上
快把酒满上干了这杯大声歌唱
好朋友好朋友今宵多欢畅
理想改变了我们的模样
也让我懂得了要珍惜朋友的肩膀
阳光总会在风雨之后撒向苍茫
让我们抬起头学会了坚强
就像朋友的这杯烈酒烫我心上
温暖我凡事别再放心上
这个吗?

经常发生小的地震,就不会发生后果严重的强烈地震.因为,通过小震已经将地下集聚的能量释放出来,减少了出现灾难性后果的情况.

设高峰期用水X,低峰Y
0.2Y=X
4X+3.2Y=44.8
解得：X=2.24;Y=11.2,共用水2.24+11.2=13.44

There are two types of vessels to go to Singapore--10-day-arrival vessel and 14-day-arrival vessel.In general,we can choose 10-day-arrival direct quick vessel.But since recently,Pan ocean shipping company has bankrupted so that the space for the whole Southeast Asian ship route is very limited.The quick vessel that we reserved before doesn't have any space,so we have to choose the 14-day-arrival vessel passing by many ports.Please kindly undertand us.

.嗯 对了,我目前上的ABC夭下口语的教师说过,事实上想征服英语是很容易的~一定有个恰当的学习情境和闇练口语对象 最关键就是外教水平,口语标准才行,不间断每日口语交流,1v1家教式教学才能有非常.好.的学习效果!课后记得回放复习课堂音频 帮助加强记忆 如果真的没人帮忙的话 最好能到听力室或BBC取得课余教材阅读 多用耳听、眼观、嘴动、脑想,很快的语境就培养起来 学习效益是必定最佳的!Good morning everybody!It's my honor to speak here,and I am very glad to share my topic with you. Then today I'd like to talk sometng about how first impression is important. It is said that first impressions are lasting impressions.peolpe form opinions and make judgement about us the first time they or hear us .And their opinions about us cannot be changed easily later.So it is important for us to creat a good first impression. Firstly, Dress neatly .we can't judge a book by its cover ,but many of us often make judgements by their appearance.so dressing neatly is necessary. Secondly, A more positive and cheerful first impression will bring us closer . Trdly,Behave ourselves. Being polite and friendly with others will help us earn their favor. it would have a lasting effect on people so it's best to smile when approacng someone, or to listen carefully when someone has sometng to . In a word, These are some of the key factors in giving others a good first impression. Thank you for listening.

我们这一代孩子呀都被外界称为 ‘温室的花朵’,就像那娇小温柔的花经不起风吹雨淋一样,而我们则是被爸爸妈妈宠坏的小公主、小皇帝一样,永远依偎在那爸爸妈妈温暖、厚重的羽翼下面,刮风了妈妈的羽翼护着我,下雨了爸爸的羽翼为我遮雨,不会有一滴雨点儿打湿我的衣服.尽管那爸爸妈妈的羽翼受了伤.
而我就是一个被爸爸妈妈宠坏的小孩,我非常依赖于他们,当他们不在我身边时,我的生活则乱如一团麻,早上起床找不到袜子；昨天做的卷子做完不知道放哪了,这生活中一切一切琐碎的事情都得依赖父母,有时我也很想摆脱对父母的依赖,但这一切都早已定形,还能改掉吗.
有时,父母也试图让我摆脱对他们的依赖,就把他们在报纸或电视上看的有关‘孩子依赖于父母’的事情讲给我听,让我印象最深刻的故事便是那次.有一天妈妈对我说她又在报纸上看到了这样一则报道,报纸上是这样说的;有一个大学生他的学习成绩等各方面都很优秀,但唯一的缺点就是太依赖于他的父母,依赖到什么可怕的程度呢,就是当他考上名牌大学时他却很忧郁,这要是换作别人肯定会高兴死的,为什么呢,就是他上大学后就要离开父母,一个人生活必须要自己照顾自己,但他真的什么都不会做,他连自己的袜子都不会洗,都是母亲在帮他洗,无奈之下,母亲与他共同搬进了学校,母亲在学校当了一个卫生管理员,一边工作一边照顾他,而他的同学呢都用异样的眼光看他.
听完这个故事后我想到自己.虽然那个男孩的学习是那么优异,但他连自己都照顾不了,又怎么照顾的了他的父母呢.
现在想起自己之前没有父母陪伴的日子,才意识到我原来是如此的依赖父母,我不想使自己成为那个男孩的影子,依赖父母到如此程度.从现在开始,我要试着并努力改变自己,不要完全依赖父母.
因为,在未来的日子,我也想用自己温暖、厚实的羽翼来保护别人.

①改“储蓄”为“储备”,改“种子”为“食物”
②改“尽管”为“不管”,改“想到”为“记住”

为什么它如此重要学好英语呢？

你认为有其他人无法得到的信息，这将是有趣吗？通话和写信有趣的人，其他人无法沟通？每当你打开你的嘴，你周围的人留下深刻印象？大的跳跃在你的职业生涯，让其他人在后面哩？

您可以得到所有这些，如果你的英语说得很好。

英语是国际语言，这意味着60％的世界人口以英语发言。这意味着，我们可以沟通好，如果我们不知道如...

Plan for Free Time休闲计划When I have free time, I would like get up early to walk in the forest in the morning, because I want to breathe the fresh air.I think it's a good way to keep healthy.在我空...

delivery in 20 days after the receival of the sight Irrevocable Letter of Credit

解题思路：先根据题意列出一次函数关系式，再根据求一次函数值的范围求自变量的取值范围．

设调进绿豆x吨，绿豆市场价格为y元/千克，
则y=16-0.01x，
依题意，8≤y≤10，即8≤16-0.01x≤10，
解得：600≤x≤800；
答：调进绿豆应在600吨到800吨之间．

点评：
本题考点： 一次函数的应用；一元一次不等式组的应用．

考点点评： 本题考查的是一次函数和一元一次不等式组在实际生活中的应用，比较简单．

Thank you for your support,i am sorry but your order will be posted to you as soon as possible .please wait a very short time.

(1)y=50+2x
(2)当定价为30时,销量为66千克,每千克利润为10元,则总利润为660元

蓝色
有点：享受敏感而有深度的交流；设身处地的体会他人；能记住谈话时共鸣的感情和思想；喜欢小群体交流的思想碰撞；关注谈话的细节.
局限：不知不觉地说教和上纲上线；原则性强,不易妥协；强烈期待别人具有敏感度和深度能理解自己；以为别人能够读懂自己的心思；不太主动与人沟通；不喜欢制造麻烦给别人,也讨厌别人制造麻烦给自己；真诚开放心胸与人互动会比较困难；习惯以防卫的状态面对别人.

灵魂战车2 英文影评:
This is a really good movie. It is fun, entertaining, and visually pleasing. The only drawback to it is that the action overshadows the story by a little too much to give it a higher rating.
The movie starts out with an intense chase with bullets flying and cars crashing and it never really lets up from there. It could have used a little more down time to build the character of the boy, Danny and explain the growing camaraderie between him and Johnny. That would make us more apt to believe Johnny was able to overcome the nature of the rider to destroy all evil enough to save the boy and his mother in the end.
At the apex of the movie is a very long, very fiery battle in mine/construction site/bad guy hangout. The viewer can tell they blew the pyrotechnics budget on this one scene. The problem is there is too much fire, too many explosions, too many bad guys getting burned, too many guns and rockets going off. It quickly becomes too much to focus on and all you can do is sit back glassy eyed and wait for it to come to its conclusion. Fortunately there isn't any dialog during that time to worry about. When I buy this on DVD I can use this scene to go to the bathroom or get coffee without hitting pause. ;-)
Central to the plot is the attempt to get the boy to a safe place. Once there we find out in short order that it isn't safe after all. In what is the biggest let down of the movie, there is not enough back story regarding the monks at "Sanctuary." We aren't given enough characterization time to get to know them so that we can feel justified or betrayed when they do what they do. I think the story must have been there at one time and was likely cut to add more action time. That is a sad thing too since the marvelous Christopher Lambert has such a short part.
Other than being action heavy, it is a very good movie. Not as good as the first one, but still very good.

“上帝粒子”只是世上的人喜欢在名称上夺人眼目,这样叫的.就和什么“美容圣经”“成功圣经”一样.根本和造物主无关.就像你的身体是由假设原子组成的,那原子又怎么解释你有思想呢?
看看他们的假说吧：137亿年前宇宙是个火球,基本粒子没有质量在宇宙间以光速乱窜,10兆分之一秒后宇宙冷却了.出现真空,这时一直在暗处潜伏着的“希玻斯”所谓上帝粒子适时的出现,填补这个空缺,给与基本粒子质量,使得宇宙带上粘性.从此以后一切不再乱窜.后来猿又变成了人.
你觉得这个解释可信吗?是不是非常牵强附会?
宇宙的奥秘已经由神的话语显明了,越读圣经越觉得无限的奥秘和爱蕴藏其中.这种爱的力量不是任何毫无生气的“粒子”可以解释的.建议多读圣经,人类需要谦卑.
借使徒保罗的话说“自从造天地以来,　神的永能和神性是明明可知的,虽是眼不能见,但借着所造之物,就可以晓得,叫人无可推诿.”

优势:劳动力比较充足.问题:人口老龄化

1
Victory still out of battling Reds' reach
Posted October 24,2009 20:43:00
Grinding it out...Hughes scored Adelaide's lone goal,but the contest was closer than the scoreline.(Getty Images:Lucas Dawson)
Map:Melbourne 3000
A gutsy second half effort by Adelaide United gave Melbourne a fright but the reigning A-League premiers escaped with a 3-1 victory at Docklands on Saturday evening.
An 88th-minute goal to Adrian Leijer,completely untroubled by the visitors' defence,was enough to seal United's fate for good after youngster Mate Dugandzic scored the opening two goals.
The three points sent Melbourne temporarily to the top of the A-League,and it will stay there if Gold Coast and Sydney lose this weekend.
Adelaide is left hovering in the middle of the competition table after 12 rounds.
The grand final rematch was the eighth time in a row the Reds have lost against Melbourne,but they often looked every bit as good as their opponents.
Coach Aurelio Vidmar said earlier in the week that the team had been working on self-belief and it certainly showed after the break,when the visitors came out with renewed vigour and kept the Victory on a tight leash.
But a strike by Adam Hughes in the 63th minute could only lessen the blow for United.
He turned a sweet header from Lloyd Owusu around the corner and into the back of the net past Glen Moss,who had a busy night in the box as Adelaide attacked with fervour.
Dugandzic banged in two goals,one on either side of the break,to send the Reds a message that they would have to battle for a win in front of a rabid Melbourne crowd.
He opened the scoring in the 17th minute off the back of an assist from Thai international Sutee Suksomkit,who started for the Victory in his first A-League match.
Sutee seemed to have plenty of confidence on the field and was given ample opportunity on the attack as he shared the midfield with Dugandzic.
Dugandzic extended Melbourne's lead when he finished from a tidy Carlos Hernandez setup after Matthew Kemp threaded a long ball through the ambling Adelaide defence.
The 20-year-old's second goal was almost a nod to the referees,who dealt him a yellow card just minutes earlier for talking back.
2
Lockyer salivating at Marshall challenge
Posted October 24,2009 19:46:00
Opposing numbers and skippers are set to go head-to-head.(Getty Images:Cameron Spencer)
The one-on-one battle between Benji Marshall and Darren Lockyer is the one that could decide the war,and it has the Kangaroos' skipper salivating ahead of his side's opening Four Nations clash against New Zealand on Sunday morning (AEDT).
While each will need a little help from team-mates,particularly the forwards,whoever comes out on top in this man-on-man match-up at The Stoop in London will put their side within sight of a Four Nations final berth.
"It's not regular that your opposite number is also the opposition captain,but that's a little challenge out there on the field," Lockyer said.
"As I start to think more about the game between now and kick-off,they're are one of the little battles that will get the blood flowing."
Marshall,too,is eagerly awaiting the showdown.
"I love challenging myself ...to get the opportunity to play against the world's best five-eighth in a Test match is something special," he said.
While both will be doing their best to unlock the other side's defence,there is little doubt they will also be called upon to do plenty of tackling.
And it will not be just any tackling - it is stopping Goliaths like Kiwi backrower Frank Pritchard and rampaging Kangaroos lock Anthony Watmough.
Australian backrower Trent Waterhouse has been handed the job of protecting Lockyer,with his main aim keeping Penrith team-mate Pritchard quiet.
"The challenge is there for Trent to come out on top of his team-mate,and I'm sure Pritchard's thinking the same thing," Kangaroos coach Tim Sheens said.
"If anyone knows [Pritchard],Trent knows him.
"It's going to be a good challenge on that side.I said to Trent that if Pritchard gets to Lockyer,then Trent's had a bad game."
The match kicks off at 6:00am AEDT Sunday morning.
都是最新的
其他的在这里：

1、B
2、D

hello,boys and girls,today I want to introduce harry potter.I believe most of the students read the harry potter movies,it is generally accepted that history's most successful film series.Premiere box-office,praise countless billions.Footage and exciting plot attracts countless fans.
But personally I think,harry potter series are greater than the film more exciting good-looking.Movie missed many exciting plot.This also no wonder that harry potter is the second,the first is the bible sales.Harry potter author jk rowling global fame,she a suit of spirals,she used to be a poor single mothers,before she often at a small cafe in the creation of the harry potter in covering card and now she valued has more than Britain's queen.

解题思路：A、根据组成“干水”的物质种类分析；
B、根据“干水”与干冰成分分析；
C、水滴是大量水分子的集合体；
D、根据干水的构成分析．

A、由题意可知，“干水”的每个颗粒内部是一颗水滴，外层是二氧化硅，故干水是水和二氧化硅组成的混合物；故选项说法正确；
B、干水和干冰不是同一种物质，干冰是固体的二氧化碳，干水是水和二氧化硅的混合物．故选项错误；
C、水是由水分子构成的，一滴水中含有1.67×10²¹个水分子，故每个颗粒的“干水”中含有大量的水分子；故选项说法错误；
D、由题意可知，“干水”的每个颗粒内部是一颗水滴，外层是二氧化硅，水与二氧化硅没有发生化学反应；故选项说法错误．
故选A．

点评：
本题考点： 纯净物和混合物的判别；物质的构成和含量分析．

考点点评： 本题属于信息给予题，难度不大，考查同学们新信息获取、处理及灵活运用所学知识进行解题的能力；理解题意、掌握物质的组成、分类等是正确解题的关键．

丛林短裤
1
这是星期五下午。戴上3班,等待他们的外套在一线回家。莱尼是在背后,因为zip在他的厚夹克(防水衣)卡住了(卡住)。他太忙于zip,他几乎错过了什么考克斯老师说。
“这儿有你的一封信所有。考克斯先生说。“别失去它,别忘了给你的父母。里面有好消息。”
莱尼想要知道这个好消息是。他在操场上跑去见他的妈妈。
“快!打开这封信,”他说。“考克斯说,有好消息里面”
“我打个招呼先!“笑他妈妈,但她打开信

解题思路：草原上不同地段上分布着种类和密度不同的牧草，体现了群落的水平结构，在生态系统中的调节作用，则为生物多样性的间接价值．生态系统的恢复力稳定性则是指生态系统遭到破坏后恢复到原状的能力．毛刺信息通过物理过程传播，故为物理信息，该信息的调节体现了调节生物的种间关系，以维持生态系统稳定的功能．

（1）草原上不同地段上分布着种类和密度不同的牧草，即水平方向上呈镶嵌分布，体现了群落的水平结构，在生态系统中的调节作用，则为生物多样性的间接价值．
（2）由于过度放牧而破坏环境，使群落演替按照不同于自然演替的方向和速度进行，恢复力稳定性则是指生态系统遭到破坏后恢复到原状的能力．
（3）动物通过观察发现毛刺，故该信息通过物理过程传播，故为物理信息，毛刺有效降低动物对黄花刺茄的取食欲望．保护了植物本身，故体现了生态系统中的信息具有调节生物的种间关系，以维持生态系统稳定的功能．
（4）假定黄花刺茄的种群初始数量为N₀，种群数量每年都以150%的增长率（出生率-死亡率）增加，则两年后它的数量为N₀×150%×150%=2.25N₀．
故答案为
（1）水平结构 间接价值
（2）速度和方向 恢复力
（3）信息传递
（4）2.25N₀

点评：
本题考点： 群落的结构特征；群落的演替；生态系统中的信息传递；生态系统的稳定性．

考点点评： 本题综合考查了生物群落和生态系统信息传递和稳定性的内容，考查了学生的理解和知识运用能力．

考试呢吧少年~

跨有荆益
兴复汉室,成就霸业
内修政理
西和诸戎,南抚夷越,外结好孙权
从荆州向宛、洛,从益州出秦川

革命题材的电影当然是以前的好,因为当时拍摄的时候肯定是有深刻意义的,你看的时候也会有比较深的感触,而现在翻拍的很多电影都肯定没有当年如此深刻,写起来就比较难了.

焚烧秸秆一是造成秸秆资源浪费,秸秆除了作为燃料、饲料、肥料外,还可用于食用菌种植、造纸和生产新型墙体材料.
二是焚烧秸秆污染了大气环境.会产生大量CO、CO2、SO2等污染物质；
三是焚烧秸秆会诱发火灾,有可能造成巨大经济损失；据相关资料报道,河南省每年因燃烧秸秆造成直接经济损失达10亿元之多.
四是焚烧秸秆使土壤表面温度增高,会烧死大量的土壤微生物,土壤水分也会损失65％－80％,使土壤板结不耐旱,吸水保墒能力大幅度下降.
五 对交通安全构成威胁.露天焚烧秸秆带来的一个最突出的问题就是焚烧过程中产生滚滚浓烟,直接影响民航、铁路、高速公路的正常运营,对交通安全构成潜在威胁.机场每逢农作物收割季节都深受秸秆露天焚烧的危害,有时机场能见度低于400米,严重影响机场航班正常起飞和降落,航空公司及旅客对此反映强烈,一旦导致事故,将造成极为不良的社会影响.

综合胜负来看,甲队和乙队都是2胜1平2负.根据惯例,双方比净胜球.甲队净胜球为1,乙队净胜球为-1.所以甲队稍占优势.双方总进球数也是甲多.但是,甲队最近三场比赛就没赢过乙队,再加上主客场的因素,双方第六场比赛的胜负很难说.
理论上,如果两队实力相当,甲队赢面大一些,毕竟甲在数据上占优.但如果考虑到足球的各种因素,我认为十分难说.有一句话叫做：“足球是圆的,什么都有可能发生.”所以楼主提供的数据根本不足以预测.

甲队
甲队比赛成绩的方差是0.24 ,乙队比赛成绩的方差是1.04
0.24﹤1.04 所以甲队成绩较稳定,甲队获胜可能性大

但愿能和自己相爱的人和亲人在一起长长久久,
即使不能在一起,也想和家乡的亲人共享这一轮明月.

小明第四次测试成绩=85-（-8）-12+10=91分

装完这批货需要B型汽车X辆,每辆B型汽车装可装Y台计算机
则由体重条件可得：A：270=3/4Y（X+1）
B：270=XY-30
楼主会解这个方程组吧,本人就不再啰嗦了.

请假条
尊敬的XX老师：
因雨天路滑,本人不小心摔倒,扭伤了脚,因此不能参加近期的体育锻炼活动和课间操,特此请假三天.恳请批准.
此致
敬礼!
请假人：张小华
时间：2011年4月25日
修改病句：因此不能参加近期的体育锻炼和课间操活动.
我改得比较多,但是一般请假条都该这么写,你可以参考一下.
那个~~~~~ 我把“请假条”三个字居中了,但是现在它自己变到了左边,“请假人”和“时间”应该在右边.

A    

（1）理解父母的有效方法是换位思考。
（2）①在家庭交往中，与父母不必太计较；②认真聆听，聆听中获得教益；③帮助父母，用行动感动亲人；④赞赏父母，赞赏中增进亲情。

B

分析：根据题目所给新信息可知，“干水”的每个颗粒内部是一颗水滴，外层是二氧化硅；据此结合所学化学知识进行分析判断即可．
A、根据题意，“干水”的每个颗粒内部是一颗水滴，外层是二氧化硅，水与二氧化硅没有发生化学反应；故选项说法错误．
B、根据题意，“干水”的每个颗粒内部是一颗水滴，外层是二氧化硅，故干水是水和二氧化硅组成的混合物；故选项说法正确．
C、水是由水分子构成的，一滴水中含有1.67×10 ²¹ 个水分子，故每个颗粒的“干水”中含有大量的水分子；故选项说法错误．
D、干水和干冰不是同一种物质，干冰是固体的二氧化碳，干水是水和二氧化硅的混合物．故选项错误．
故选B．

以下仅供参考：
2007年内部销售商品未实现利润=40×5-40×3=80
2007年内部销售固定资产未实现利润=原价中含有的未实现损益（100-60）-多计提的折旧（10-6）=36
2007年内部销售未实现利润=80+36=116
2008年内部销售商品未实现利润=20×6-20×3=60
2008年内部销售固定资产未实现利润=36-（20-12）=28
2008年内部销售未实现利润=60+28=88

要点：连日来，单日微博关注“光盘行动”的人数不仅大幅上升，各路媒体也纷纷予以关注。

众数是3.8米，4.0米；中位数是3.9米；极差是0.6米；平均数是3.9米．
故选C．

冬季,从蒙古-西伯利亚高原吹来的西北季风干冷,经过温暖的日本海域带来大量的水汽,到达日本后受到地形抬升形成降雨,降雨受冷空气影响形成降雪.

The factory is suffering a power failure for 3 or 4 days due to the recent typhoon, as a result, the goods may not be ready on time. We will try to send them to you one after another for your cofirmation. Sorry for any inconvenience.

1、B
2、D

A

解题思路：此题考查传染病的预防措施：控制传染源、切断传播途径、保护易感人群，据此答题．

传染病能够在人群中流行，必须具备传染源、传播途径、易感人群这三个环节，缺少其中任何一个环节，传染病就流行不起来，传染源是指能够散播病原体的人或动物；传播途径是指病原体离开传染源到达健康人所经过的途径；易感人群是指对某种病原体缺乏免疫力而容易感染该病的人群，预防传染病的一般措施有控制传染源、切断传播途径、保护易感人群，病原体是指引起传染病的细菌、病毒和寄生虫等生物，专家号召大家搞好个人饮食卫生，勤洗手属于切断传播途径．
故选：B

点评：
本题考点： 传染病的预防措施．

考点点评： 对于传染病的知识，是中考的重点，多以选择题或是材料题的形式出现，要灵活掌握．

解题思路：（1）利用总套数1000减去其它类型的房子的套数，即可求解；
（2）按房屋的面积从小到大，第500和第501套的平均数就是中位数，据此即可确定；
（3）根据统计图，确定哪种类型的房子的套数最多，即可．

（1）1 000-480-45-30=350（套）．


（2）中位数落在80～100平方米；

（3）多建筑面积80～100平方米的住房．

点评：
本题考点： 频数（率）分布直方图；中位数．

考点点评： 本题考查读频数分布直方图的能力和利用统计图获取信息的能力；利用统计图获取信息时，必须认真观察、分析、研究统计图，才能作出正确的判断和解决问题．

Hello, everybody ! Welcome to our museum. Now you are requested to do the following things. First smoking is forbidden in the museum. Second you should keep silent in the museum . Third you should make the museum clean. Fourth you mustn't take photos in it . Fifth you can consult the guides if you want to get more information about the exibition. Finally you mustn't touch anything in the museum. The museum opens from 8:30 am to 4:30 pm during Sunday to Friday.Thank you all!

The prices of raw materials are continuesly increasing recently.

都有可能
你可以多做几个对照,如空载体转化、空细胞涂板等等,帮助缩问题小范围
你可以询问实验室其他人员,看看酶是否有问题,感受态是否有问题,抗生素是否有问题,等等
按你给的线索,菌落很少,很有可能是切开的线性质粒没有连上片段,转不进细胞,少量没有切开的空载体转进细胞,形成阴性菌落.问题可能在片段酶切上,建议连T载体再切,或双酶切换成两步单酶切,或换酶切位点,或重新设计保护碱基