PCR仪使用说明

PCR仪使用说明
步骤：
1、开机：打开开关，视窗上显示“SELF TEST”，显示10秒中后，显示RUN-ENTER菜单：“-RUN
ENTER PROGRAM PROGRAM”准备执行程序。

2、放入样本管，关紧盖子。

3、如果要运行已经编好的程序，则直接按《Proceed》，用箭头键选择已储存的程序，按《Proceed》，
则屏幕显示：“-ENABLE DISABLE HEATED LID” 按《Proceed》选择ENABLE，则开始执行程序。

4、如果要输入新的程序，则在RUN-ENTER菜单上用箭头键选择ENTER PROGRAM，按《Proceed》，
屏幕显示“-NEW LIST EDIT DELET”，按《Proceed》，1）选择NEW，命名新的程序，最多8个字母，输入后按《Proceed》确认。

2）输入程序步骤：名字输入后，显示“STEP1 _TEMP GOTO OPITON END”，按《Proceed》则可以输入温度（0～100℃），按《Proceed》确认后，则可以输入孵育时间，用《Select》键移动光标，输入数字，完成后按《Proceed》确认，跳到下一步，输入方式同上。

3）选择GOTO，输入循环步骤时链接到第几步（循环数最多可达9999次）（为实际循环数-1)。

4) 选择option，显示“STEP EXTENDI NCREMENT SLOPE”再选择increment，按《Proceed》
确认，输入初始的温度，确认后输入时间，按《Proceed》确认，然后输入每个循环增加或减少的温度，增加用正值，减少用负值（-0.1℃～6℃），按《Proceed》确认。

选择extend，按《Proceed》确认，输入每个循环增加或减少的时间（-60～60秒），按《Proceed》确认。

选择slop（指温度上升或下降的速率），输入温度的改变值（-0.1～1.5℃）按《Proceed》确认，然后输入加热或致冷的速度，按《Proceed》确认。

4）选择End，输入结束步骤。

5、输入完成的程序后，到RUN-ENTER菜单，选择新程序，开始运行。

6、其它：用《pause》可以暂停一个运行的程序，再按一次继续程序。

用《stop》或《Cancel》
可停止运行的程序。

编辑程序：
1、可以用《Cancel》键删除输错的值，输入新的值后按《Proceed》确认。

2、对于未输完的程序，要先输入END，按《Proceed》将程序储存后才能删除。

3、删除已经储存的程序：从RUN-ENTER菜单中选择RUN－PROGRAM，按《Proceed》，显示主菜单，
选择DELET，用《Select》选择删除。

4、查看程序的步骤：在主菜单上选择LIST，按《Proceed》，用《Select》将选择名称，再按《Proceed》，显示程序的第一步，用《Select》键向前、向后翻页查看，此时不能改变程序的值。

编辑已有的程序：
1、从RUN-ENTER菜单中选择RUN－PROGRAM，按《Proceed》，显示主菜单，选择EDIT，按《Proceed》确认，用《Select》键选择要编辑的程序，按《Proceed》显示程序的第一步。

2、用《Select》键将光标移到要改变的温度或循环的值上，键入新值，按《Proceed》确认，按《Cancel》删除键入的值，出现空格，键入新值后确认。

注：一旦值被改变或删除，原来的值不能恢复，必须重新键入。

3、编辑时间值：必须重新键入小时、分钟、秒，按《Proceed》。

如何设置一个PCR体系：
一般分为8步：
1、预变性：可用94～95℃，2～10min，一般用5min。

2、变性：一般用94℃，30s～2min，一般45s～1min。

3、退火：温度自定，30s～2min。

4、延伸：70～75℃，一般72℃，对于＜2kb，＜1min；＞2kb，每增加1kb加1min。

5、循环数：一般25～35个循环。

6、最终延伸：72℃，5～15min。

7、保存：10℃，时间设为0。

8、END。

PCR使用说明
来源: 实验中心作者: 实验中心发表时间: 2009-04-27
1．开机预热30 分钟
2．利用上下左右键进入到程序主页面.
3．按A 对应的list,然后利用上下键选择empty ,再按enter 键进入
4．利用A 对应的abc 和数字键进行设定程序的名称,输入完按C 对应的name ok
5．再输入顶盖预热温度如105 度,按enter 键进入即可设定每一步的温度时间,循环数及梯度6．温度的设定:直接按数字键
7．时间的设定如下:如4 小时输入4..(注意一定要输入4 后的两个点)
8． 4 分钟输入.4.
9． 4 秒直接输入4 即可
10．循环的设定:在←对应的下面输入从第几步循环,在#下输入循环数,如从第2 步到第4 步需
30 个循环,在第4 步的←应处输入数字2,在# 下输入29
11．梯度的设定:在gradient 对应下输入梯度的范围即可,如需要温度在55-65 度,在temp对应处输入60,在gradient 输入10 即可可以利用A 看已设的各个对应位置的温度.
12．设定完这些按pgmOK,再按start 程序开始运行.注意:顶盖温度较高,在使用过程中注意不要触摸用完要停下,不要让仪器长时间处于4 度,容易损坏仪器.
3V乙肝病毒PCR检测试剂盒使用说明
（20人份）
简介
乙型肝炎是一种严重危害人类健康的传染病，由乙型肝炎病毒（Hepatitis B Virus）所致，采用ELISA检测乙肝两对半的方法已广泛应用，对乙肝诊断提供了一种简便快速的工具，但由于免疫学检测只能提供HBV存在的间接证据，依靠乙肝两对半来判断病毒的复制状态及血清的传染性就受到了限制。

近年来发展起来的聚合酶链反应（PCR）技术可直接检测病毒的病原体核酸，其特异性强、敏感度高，因此PCR在临床上可作为乙肝病毒检测的有效方法，可直接反映乙肝病毒存在与否，特别适应于早期诊断及明确诊断HBC的复制和血清的传染性。

本试剂盒扩增靶序列选自HBV基因组相对保守的C区，扩增片段为270bp，具有良好的特异性和敏感度。

试剂盒组成：
裂解液A0.5（20ul/管）液体石蜡600ul
反应液110ul（5ul/管）50×TAE电泳缓冲液20ml
灭菌双蒸水1ml琼脂糖 1.0g
阳性对照50ul（5ul/管）溴乙锭（EB）20ul
实验操作
1.样品处理：
（1）标本来源：新鲜血清或血浆。

（2）标本保存：当天不处理的标本置-20℃保存。

（3）标本处理：取20ul裂解液A于离心管中，加40ul待测血清，混匀后，100℃沸水浴15分钟，15000rpm离心20分钟，4℃存放备用。

2.扩增反应
（1）根据扩增样品数量，取PCR反应液按每管5ul分装，每管再加入15ul灭菌双蒸水和一滴液体石蜡油（约20ul），盖上管盖备用。

（2）先将灭菌双蒸水5ul加入一个分装管中（作为阴性对照）。

取各标本离心上清液5ul加入对应的反应管中，最后取阳性对照5ul加入一个反应管中。

加样完毕的反应管混匀后，10000rpm离心5秒，取出放入扩增仪里。

（3）按下列步骤循环
94℃5分钟
94℃30秒
55℃30秒35次循环
72℃30秒
72℃5分钟
3.结果判断
（1）将50×TAE电泳缓冲液20ml加蒸馏水稀释至1000ml（1×TAE）待用，取50ml1×TAE缓冲液于三角瓶中，加入5ul溴乙锭，再加入1.0可琼脂糖，加热使之溶解至清亮状。

待琼脂糖凉至50℃左右时，倒入事先准备好的电泳平板上。

电泳槽中倒入1×TAE缓冲液。

（2）取15ul扩增产物，加入2%琼脂糖凝胶孔中进行电泳，80V电压电泳30分钟后，取出进行观察（至蓝色指示剂离点样孔1.5cm左右时）。

（3）将凝胶置于紫外透射仪上，在紫外灯下被溴化乙锭染色的DNA呈现明亮的橙黄色的荧光。

样品出现和阳性对照在同一水平位置上的橙黄色荧光带即为阳性，否则为阴性。

注意事项：
1.阳性对照无需进行样品处理，直接取5ul进行PCR扩增。

2.使用前请将各管试剂复温至室温，并高速离心数秒，以免粘壁损失。

3.扩增仪类型的差异和循环温度的差异，可能导致二聚体（在蓝色指示剂前面）的出现。

4.试剂盒存放于-20℃，有效期1年。

5.每次实验需设置阴、阳性对照。

6.取样品上清时，避免取出沉淀物。

7.因反应液为浓品，取样需准确，以免影响实验结果。

简单说明：
Real Time PCR反应液配制（50μl反应体系，染料法）：
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使用准备：
引物溶解：先12000rpm离心5分钟，然後用ddH2O溶解成25μM，加水量根据引物合成报告单提供的计算方法计算。

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配制说明：
按照以下配方，如果推荐引物加样量如加0.2ul的结果不好，可以调整引物使用量。

括号里是第二备选加样量和第三备选加样量。

另外，可以调整的参数就是扩增程序的退火温度，请参考说明书。

可以用小体积的，等比例缩小体系。

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提高数据重复性的小技巧：
配制PCR 反应液时，尽量采用预混再分装的方式。

例如，可以计算加同一对引物的标本为5个，准备做三复孔，采用15ul体系，引物浓度为200nM(按照上面表格是加0.4ul的），就可以用以下的加样方式：
（比5份稍微多一点），总共为130ul的2*PCR master mix，2.1ul的上游引物，2.1 ul的下游引物，加dd水至235ul；混匀分装至每管45ul，每管加待检测样品5ul，混匀平均分装成三管，每管15ul。

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加样的注意事项：
1、注意加样枪的使用，加样时不要把枪头伸入液面下，避免挂液误差；
2、要使用低吸附的进口枪头，避免每次的残留导致的加样误差。

3、正确的加样方式：离液面1-2mm，加液，不靠壁，不贴液面。

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混匀平均分装的注意事项：
1、第一管吸液前需要先把枪头在液面中浸润一下，以保证三复孔的枪头吸液量是差不多的。

2、用混匀器进行混匀，不要用移液器吹打。

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详细说明书
transhold cat. A2010A0112
荧光定量PCR Premix试剂盒(荧光染料)
（FQ-PCR PreMix Kit）(除LIGHTCYCLER)操作说明书
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【前言】
本试剂盒提供了进行简便、灵敏的PCR（或荧光PCR）检测的完整系统，包含了经过优化的缓冲液、dNTP、热启动Taq DNA 聚合酶混合物、MgCl2溶液和预混的SYBR GREEN染料，成为一个即时可用的混合物。

您不必再花费时间去混合试剂，同时确保了结果的一致性和可重复性。

使用定量PCR Premix试剂可进行实时荧光PCR检测，获得方便、灵敏和准确、可靠的检测结果。

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【规格】
50人份
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【试剂盒组成】
定量PCR Premix试剂混合物(2X): 1.25 ml
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【反应液配制】
（1）反应体系中，引物常规终浓度可设为200nM。

反应性能较差时，可以在0.05～0.9 μM（50－900nM）范围内调整引物浓度。

（2）在25 μl反应体系中，DNA模板的添加量通常在100ng以下。

因不同种类的DNA模板中含有的靶基因的拷贝数不同，必要时可进行梯度稀释，确定最佳的DNA模板添加量。

如果欲使用本制品进行2 Step RT-PCR反应的第二步PCR扩增反应，第一步的RT反应液作为DNA 模板时的添加量不要超过PCR反应液总体积的10%。

（3）适用的荧光定量PCR仪：ABI，MJ RESEARCH,BIORAD,STRATAGENE等，ROCHE品牌仪器由于光路不同，可能无法得到满意结果。

[编辑]
【扩增条件】
以ABI7000为例，Detector设为SYBR,QUENCHER设为NONE,
1、当引物TM值≥60℃时，推荐：95℃ 15分钟→95℃ 15秒→60℃ 1分钟℃→溶解曲线分析
40～50个循环（可选）
2、当引物TM值<60℃时，推荐：95℃ 15分钟→95℃ 15秒→55℃ 30秒→72℃ 45秒→溶解曲线分析
40～50个循环（可选）
用户可根据自己的用途和引物、探针设计的具体情况来确定适合的退火温度，退火时间和循环次数。

[编辑]
【使用注意事项】
1、冻结制品融解後请上下颠倒轻轻均匀混合，避免振荡器等剧烈搅拌。

混匀制品後请用离心机轻微离心後使用。

应在使用前应充分融化，混匀，离心数秒，使液体集中在离心管底部。

2、本制品使用前请于-20℃保存，融解後的制品请于冰上放置，使用後请立即保存于-20℃。

3、尽量避免多次反复冻融，如果需要多次使用时，请在第一次融解後将制品按适当量分装後保存。

4、反应液的配制、分装请一定使用新的（无污染的）枪头、Microtube等，尽量避免污染。

5、引物设计过程中尽量避免发夹结构、引物二聚体结构的发生。

目标片断最好在250bp以内，尽可能在150bp以内。

6、反应体系中，引物常规终浓度可设为200nM（上游引物200nM，下游引物200 nM）。

根据您的保存浓度不同，计算加入的引物量，使体系的终浓度保持相同。

引物浓度的计算方法可以参考：《荧光定量PCR实验指南》。

必要时您也可以根据体系的不同，来优化引物浓度。

反应性能较差时，可以在0.05～0.9 μM（50－900nM）范围内调整引物浓度。

7、如果使用其他体积的体系（10-50μl），应保证终浓度与说明书上的终浓度一致。

8、在操作中，应使用不含荧光物质的一次性手套(经常替换)、一次性薄壁进口200μl PCR管、移液器头(带滤嘴自卸式)，不能用手直接触摸PCR反应管。

9、操作台、移液器、离心机、扩增仪等仪器设备应经常用10%次氯酸钠及/或70%乙醇擦拭消毒。

实验房间、超净工作台应定期及每次实验後用紫外灯处理。

10、试剂盒请在保质期内使用。

11、配制反应体系过程中若需标记，请在试管架或离心机管架上标记，不要直接标记在离心管上。

12、实验中涉及的有害有毒的标本及试剂应妥善放置，专人保管；废弃物应置专门容器中，妥善处理。

13、配制反应体系时，应注意移液器的使用方法，所有的液体都要缓慢加至管底，不要加至管壁，所有液体的混匀要用振荡器进行，不能用移液器吹打，避免产生气泡。

如果产生气泡，请用手指将气泡弹破，再低速离心。

反应体系配制完毕後低速离心数秒，立即进行PCR扩增。

虽然PCR基因扩增仪的生产厂家、型号很多，工作原理不尽相同，使用方法也不尽一致，但都具有向自动化和智能化发展的趋势，这对于使用者来说比较方便，只要参考说明书轻轻触动键钮，输入或改变扩增程序、反应时间、温度等，置入扩增样品，扩增仪就会自动完成整个扩增过程。

这里仅以MJ公司的PTC－150－16型PCR仪为例，简单介绍该型机的使用和维护要点。

1. PCT-150-16型PCR仪主要技术特点
(1) 温度范围-9－105℃；
(2) 恒温精度±0.4℃；
(3) 温度均匀性12秒内样品孔温度升至92℃，孔间温差±0.4℃；
40秒内样品孔温度升至92℃，孔间温差±0.3℃。

(4) 样品容量0.5ml管×16支；
(5) 电源100－240V AC，50－60Hz，最大功率150W；
(6) 内存80个扩增程序；
(7) 规格体积，20×28×11cm，重量，3.0kg；
(8) 使用环境温度4－32℃，相对湿度10％－90％；
(9) 使用寿命正常情况下10年。

2. 使用方法
(1) 准备好反应管；
(2) 打开机盖，平稳、端正地置入反应管，盖好机盖；
(3) 打开电源开关，机器自检屏幕显示的提示设定程序。

用Proceed键起动扩增，结束时出现“Complete”，待风机停止工作，关闭电源，取出样品盖好扩增仪护套。

3. 维护注意事项
(1) 注意本机的使用环境条件和电源；
(2) 机盖开关要轻，以防损坏盖锁；
(3) 严禁工作时打开机盖；
(4) 定期用中性肥皂水清洗样品槽，严禁使用强碱、有机溶液和高浓度酒精擦洗；
(5) 有故障时请有专业知识的维修人员或厂家技术人员维修。

（英文版）
Two regulations promulgated for implementation is in the party in power for a long time and the rule of law conditions, the implementation of comprehensive strictly strategic plan, implementation in accordance with the rules and discipline to manage the party, strengthen inner-party supervision of major initiatives. The two regulations supporting each other, the < code > adhere to a positive advocate, focusing on morality is of Party members and Party leading cadres can see, enough to get a high standard; < rule > around the party discipline, disciplinary ruler requirements, listed as "negative list, focusing on vertical gauge, draw the party organizations and Party members do not touch the" bottom line ". Here, the main from four square face two party rules of interpretation: the first part introduces two party Revised regulations the necessity and the revision process; the second part is the interpretation of the two fundamental principles of the revision of laws and regulations in the party; the third part introduces two party regulations modified the main changes and needs to grasp several key problems; the fourth part on how to grasp the implementation of the two regulations of the party. < code > and < Regulations > revised the necessity and revised history of the CPC Central Committee the amendment to the Chinese Communist
Party members and leading cadres honest politics several guidelines > and < Chinese Communist Party discipline and Punishment Regulations > column 1 by 2015 to strengthen party laws and regulations focus. Two party regulations revision work lasted a Years, pooling the wisdom of the whole party, ideological consensus, draw historical experience, respect for the wisdom of our predecessors, which reflects the unity of inheritance and innovation; follow the correct direction, grasp the limited goals, adhere to the party's leadership, to solve the masses of the people reflect a focus on the problem. The new revision of the < code > and < rule >, reflects the party's 18 and the eighth session of the third, the spirit of the fourth plenary session, reflecting the experience of studying and implementing the General Secretary Xi Jinping series of important speech, reflects the party's eighteen years comprehensive strictly practice. (a) revised two regulations of the party need of < the ICAC guidelines > in < in 1997 Leaders as members of the Communist Party of China clean politics certain criteria (Trial) > based on revised, the promulgation and implementation of January 2010, to strengthen the construction of the contingent of leading cadres play an important role. But with the party to manage the party strictly administering the deepening, has not been able to fully meet the actual needs. Content is too
complicated, "eight prohibition, 52 are not allowed to" hard to remember, and also difficult to put into practice; the second is concisely positive advocated by the lack of prohibited provisions excessive, no autonomy requirements; the third is banned terms and discipline law, both with the party discipline, disciplinary regulations repeat and Criminal law and other laws and regulations repeat; the fourth is to "clean" the theme is not prominent, not for the existing problems, and is narrow, only needle of county-level leading cadres above. < rule > is in 1997 < Chinese Communist Party disciplinary cases (Trial) > based on revision, in December 2003 the promulgation and implementation, to strengthen the construction of the party play very important role. Along with the development of the situation, which many provisions have been unable to fully meet the comprehensive strictly administering the practice needs. One is Ji law, more than half of the provisions and criminal law and other countries laws and regulations Repetition; two is the political discipline regulations is not prominent, not specific, for violation of the party constitution, damage the authority of Party Constitution of misconduct lack necessary and serious responsibility to pursue; third is the main discipline for the leading cadres, does not cover all Party members. Based on the above situation, need to < the criterion of a clean and honest
administration > and < rule > the two is likely to be more relevant regulations first amendment. By revising, really put the authority of Party discipline, the seriousness in the party tree and call up the majority of Party members and cadres of the party constitution of party compasses party consciousness. (II) two party regulations revision process the Central Committee of the Communist Party of China attaches great importance to two regulations revision . Xi Jinping, general books recorded in the Fifth Plenary Session of the eighth session of the Central Commission for Discipline Inspection, on the revised regulations < > made clear instructions. According to the central deployment, the Central Commission for Discipline Inspection from 2014 under six months begin study two regulations revision. The Standing Committee of the Central Commission for Discipline Inspection 4 review revised. Comrade Wang Qishan 14 times held a special meeting to study two regulations revision, amendment clarifies the direction, major issues of principle, path and target, respectively held a forum will listen to part of the province (area) secretary of the Party committee, Secretary of the Discipline Inspection Commission, part of the central ministries and state organs Department
The first party committee is mainly responsible for people, views of experts and scholars and grassroots party organizations and
Party members. Approved by the Central Committee of the Communist Party of China, on 7 September 2015, the general office of the Central Committee of the Party issued a notice to solicit the provinces (autonomous regions, municipalities) Party, the central ministries and commissions, state ministries and commissions of the Party (party), the General Political Department of the military, every 3 people organization of Party of two regulations revision opinion. Central Commission for Discipline Inspection of extensive solicitation of opinions, careful study, attracting, formed a revised sent reviewers. In October 8 and October 12, Central Committee Political Bureau Standing Committee and the Political Bureau of the Central Committee After consideration of the two regulations revised draft. On October 18, the Central Committee of the Communist Party of China formally issued two regulations. Can say, two laws amendment concentrated the wisdom of the whole party, embodies the party. Second, < code > and < Regulations > revision of the basic principles of two party regulations revision work and implement the party's eighteen, ten eight plenary, the spirit of the Fourth Plenary Session of the Eleventh Central Committee and General Secretary Xi Jinping important instructions on the revised < low political criterion > and < Regulations >, highlighting the ruling party
characteristics, serious discipline, the discipline quite in front of the law, based on the current, a long-term, advance as a whole, with Bu Xiuding independent < rule > and < rule >. Main principle is: first, adhere to the party constitution to follow. The constitution about discipline and self-discipline required specific, awaken the party constitution of party compasses party consciousness, maintaining the authority of the constitution. General Secretary Xi Jinping pointed out that "no rules, no side round. Party constitution is the fundamental law, the party must follow the general rules. In early 2015 held the eighth session of the Central Commission for Discipline Inspection Fifth Plenary Session of the 16th Central Committee, Xi Jinping again pointed out that constitution is the party must follow the general rules, but also the general rules." the revision of the < code > and < rule > is Method in adhere to the regulations established for the purpose of combining rule of virtue is to adhere to the party constitution as a fundamental to follow, the constitution authority set up, wake up the party constitution and party rules the sense of discipline, the party constitution about discipline and self-discipline specific requirements. 4 second is to adhere to in accordance with the regulations governing the party and the party. The Party of rule of virtue "de", mainly refers to the party's ideals and beliefs, excellent traditional style. The revised the
< code > closely linked to the "self-discipline", insisting on the positive initiative, for all members, highlight the "vital few", emphasized self-discipline, focusing on the morality, and the majority of Party members and the ideological and moral standards. The revised < > Ji method separately, Ji, Ji Yan to Method, as a "negative list", emphasizing the heteronomy, focusing on vertical gauge. Is this one high and one low, a positive reaction, the strict party discipline and practice results transformation for the integration of the whole party to observe moral and discipline requirements, for the majority of Party members and cadres provides benchmarking and ruler. Third, insist on to. In view of the problems existing in the party at the present stage, the main problems of Party members and cadres in the aspect of self-discipline and abide by the discipline to make clearly defined, especially the party's eighteen years strict political discipline and political rules, organization and discipline and to implement the central eight provisions of the spirit against the four winds and other requirements into Disciplinary provisions. Not one pace reachs the designated position, focusing on in line with reality, pragmatic and effective. After the revision of major changes, major changes in the < code > and < rule > modified and needs to grasp several key problems (a) < code > < code > adhere to according to regulations
governing the party and party with morals in combination, for at the present stage, the leadership of the party members and cadres and Party members in existing main problems of self-discipline, put forward principles, requirements and specifications, showing Communists noble moral pursuit, reflected at all times and in all over the world ethics from high from low 5 common requirements. One is closely linked to the "self-discipline", removal and no direct relation to the provisions of . the second is adhere to a positive advocate, "eight prohibition" 52 are not allowed to "about the content of the" negative list moved into synchronization amendment < cases >. Three is for all the party members, will apply object from the leadership of the party members and cadres to expand to all Party members, fully embodies the comprehensive strictly required. The fourth is prominent key minority, seize the leadership of the party members and cadres is the key, and put forward higher requirements than the ordinary Party members. Five is to simplify, and strive to achieve concise, easy to understand, easy to remember. The revised < code > is the ruling Party since the first insists on a positive advocate forAll Party members and the self-discipline norms, moral declaration issued to all members of the party and the National People's solemn commitment. > < criterion of a clean and honest administration consists of 4 parts, 18,
more than 3600 words. After the revision of the < code >, a total of eight, 281 words, including lead, specification and Party member cadre clean fingered self-discipline norms, etc. Part 3 members low-cost clean and self-discipline, the main contents can be summarized as "four must" "eight code". Lead part, reiterated on ideal and faith, fundamental purpose, the fine traditions and work style, noble sentiments, such as "four must" the principle of requirements, strong tone of self-discipline, The higher request for 6 and supervised tenet, the foothold in permanent Bao the party's advanced nature and purity, to reflect the revised standards requirements. Members of self-discipline norms around the party members how to correctly treat and deal with the "public and private", "cheap and rot" thrifty and extravagance "bitter music", put forward the "four norms". Party leader cadre clean fingered self-discipline norms for the leadership of the party members and cadres of the "vital few", around the "clean politics", from civil servant of the color, the exercise of power, moral integrity, a good family tradition and other aspects of the leadership of the party members and cadres of the "four norms" < > < norm norm. "The Party member's self-discipline norms" and "party members and leading cadre clean fingered self-discipline norms," a total of eight, collectively referred to as the "eight". "Four must" and "eight" of the
content from the party constitution and Party's several generation of leaders, especially Xi Jinping, general secretary of the important discussion, refer to the "three discipline and eight points for attention" statements, and reference some embody the Chinese nation excellent traditional culture essence of epigrams. (2) the revised regulations, the main changes in the revised Regulations > to fully adapt to the strictly requirements, reflects the according to the regulations governing the law of recognition of deepening, the realization of the discipline construction and Jin Ju. < rule > is party a ruler, members of the basic line and follow. And the majority of Party members and cadres of Party organizations at all levels should adhere to the bottom line of thinking, fear discipline, hold the bottom line, as a preventive measure, to keep the party's advanced nature and purity. 1, respect for the constitution, refinement and discipline. Revised < rule > from comprehensive comb physical constitution began, the party constitution and other regulations of the Party of Party organizations and Party discipline requirements refinement, clearly defined in violation of the party constitution will be in accordance with regulations to give the corresponding disciplinary action. The original 10 categories of misconduct, integration specification for political discipline, discipline, honesty and discipline masses Ji Law and discipline and discipline and
other six categories, the content of < rule > real return to Party discipline, for the majority of Party members and listed a "negative list. 7 2, highlighting the political discipline and political rules. > < Regulations according to the stage of the discipline of outstanding performance, emphasizing political discipline and political rules, organization and discipline, in opposition to the party's leadership and the party's basic theory, basic line, basic program and basic experience, the basic requirement of behavior made prescribed punishment, increase the cliques, against the organization such as violation of the provisions, to ensure that the central government decrees and the Party of centralized and unified. 3, adhere to strict discipline in the law and discipline In front, Ji separated. Revised < Regulations > adhere to the problem oriented, do Ji separated. Any national law existing content, will not repeat the provisions, the total removal of 79 and criminal law, repeat the content of the public security management punishment law, and other laws and regulations. In the general reiterated that party organizations and Party members must conscientiously accept the party's discipline, die van comply with national laws and regulations; at the same time, to investigate violations of Party members and even criminal behavior of Party discipline and responsibility, > < Regulations distinguish five different conditions, with special provisions were
made provisions, so as to realize the connection of Party discipline and state law. 4, reflect Wind building and anti-corruption struggle of the latest achievements. < rule > the party's eighteen years implement the spirit of the central provisions of the eight, against the requirements of the "four winds" and transformation for disciplinary provisions, reflecting the style construction is always on the road, not a gust of wind. In the fight against corruption out of new problems, increase the trading rights, the use of authority relatives profit and other disciplinary terms. Prominent discipline of the masses, the new against the interests of the masses and ignore the demands of the masses and other disciplinary terms and make provisions of the disposition and the destruction of the party's close ties with the masses.
Discipline to protect the party's purpose. 8 of these regulations, a total of three series, Chapter 15, 178, more than 24000 words, after the revision of the regulations a total of 3 series, Chapter 11, 133, 17000 words, divided into "general" and "special provisions" and "Supplementary Provisions" Part 3. Among them, add, delete, modify the provisions of the proportion of up to nearly 90%. 1, the general general is divided into five chapters. The first chapter to the regulations of the guiding ideology, principles and scope of application of the provisions, highlight the strengthening of the party
constitution consciousness, maintenance the authority of Party Constitution, increase the party organizations and Party members must abide by the party constitution, Yan Centralized centralized, would examine at all levels of the amended provisions implementing and maintaining Party discipline, and consciously accept the party discipline, exemplary compliance with national laws and regulations. The second chapter of discipline concept, disciplinary action types and effects of the regulations, will be a serious warning from the original a year for a year and a half; increase the Party Congress representative, by leaving the party above (including leave probation) punishment, the party organization should be terminated its representative qualification provisions. The third chapter of the disciplinary rules of use prescribed in the discipline rectifying process, non convergence, not close hand classified as severely or heavier punishment. "Discipline straighten "At least eighteen years of five years, these five years is to pay close attention to the provisions of the central eight implementation and anti -" four winds ". The fourth chapter on suspicion of illegal party disciplinary distinguish five different conditions, with special provisions were made provisions, to achieve effective convergence of Party and country 9 method. < rule > the provisions of Article 27, Party organizations in the
disciplinary review found that party members have committed embezzlement, bribery, dereliction of duty dereliction of duty and other criminal law act is suspected of committing a crime shall give cancel party posts, probation or expelled from the party. The second is < Regulations > Article 28 the provisions of Party organizations in the disciplinary review But found that party members are stipulated in the criminal law, although not involved in a crime shall be investigated for Party discipline and responsibility should be depending on the specific circumstances shall be given a warning until expelled punishment. This situation and a difference is that the former regulation behavior has been suspected of a crime, the feeling is quite strict, and the latter for the behavior not involving crime, only the objective performance of the provisions of the criminal code of behavior, but the plot is a crime to slightly. < Regulations > the 29 provisions, Party organizations in the discipline review found that party members and other illegal behavior, affect the party's image, the damage to the party, the state and the people's interests, we should depend on the situation Seriousness given disciplinary action. The loss of Party members, seriously damaging the party's image of behavior, should be given expelled from the party. At this article is party member is in violation of the criminal law outside the other illegal acts, such as violates。