ColonyPCR
Colony PCRDavid AmbergThis procedure will work for both yeast and E. coli:Take a small colony and suspend it in 5ul of H2O in a PCR tube. Heat for 5 min at 95?C and then spin the condensation down in a microfuge. Set up the PCR reaction as follows: 5ul H2O + cells5ul 5uM primer25ul 10XTaq Buffer5ul 2mM dNTPs0.5ul 10 mg/ml acetylated BSA1ul Taq DNA polymerase23.5 ul H2OPCR Conditions: 94?C x 4......閱讀全文
Colony－PCR
Colony?PCRDavid AmbergThis procedure will work for both yeast and E. coli:Take a small colony and suspend it in 5ul of H2O in a PCR tube. Heat for 5 m
Colony－PCR
Colony PCR is useful in determining whether or not a specific colony on a plate has a sequence you desire. Primers for the specific sequence should be
Colony－PCR－Protocol
1. Pull out eight glycerol stock plates from the –80oC freezer and set on bench top to thaw. Be sure to remove the foil seal before leaving the plates
Silver:－Colony－PCR
Zymolyase Solution:Regular stock of zymolyase is 10 mg/ml diluted in water, mix by inverting, not all will go into solution (filter it) store aliquots
Bacterial－Colony－PCR
Bacterial Colony?PCRObjective:This protocol allows rapid detection of transformation success when primers are available to allow determination of corr
菌落PCR（Colony－PCR）方法
菌落PCR（Colony PCR）可不必提取基因組DNA，不必酶切鑒定，而是直接以菌體熱解后暴露的DNA為模板進行PCR擴增，省時少力。建議使用載體上的通用引物。通常利用此方法進行重組體的篩選或者DNA測序分析。最后的PCR產物大小是載體通用引物之間的插入片斷大小。具體方法：1、PCR混合物的制備T
Endy:Yeast－Colony－PCR
MethodUsing sterile pipette tips, transfer a 1 mm colony into 50 uL of 60 U/ml Zymolyase3 uL of 1 U/mL Zymolyase stock solution47 uL of waterIncubate
Blackburn:Yeast－Colony－PCR
OverviewThis is a quick and easy yeast colony PCR protocol that does not require zymolyase step.Updated Protocol:?Blackburn Lab: Quick and Easy Yeast
菌落PCR（Colony－PCR）具體方法
菌落PCR（Colony PCR）可不必提取基因組DNA，不必酶切鑒定，而是直接以菌體熱解后暴露的DNA為模板進行PCR擴增，省時少力。建議使用載體上的通用引物。通常利用此方法進行重組體的篩選或者DNA測序分析。最后的PCR產物大小是載體通用引物之間的插入片斷大小。具體方法：1、PCR混合物的制
Engineering－BioBrick－vectors－from－BioBrick－parts/Colony－PCR
MaterialsPCR SuperMix High FidelityVF2 primer (5''-TGCCACCTGACGTCTAAGAA-3'')VR primer (5''-ATTACCGCCTTTGAGTGAGC-3'')De
Colony－Hybridization
ProcedurePrepare serial 10-fold dilutions of transformed bacteria in LB and spread 100 μL onto LB/Amp plates, as described in the Electroporation prot
COLONY－HYBRIDIZATION
COLONY HYBRIDIZATION1) CUT 4 PIECES OF 3MM WHATMAN PAPER AND PLACE EACH ONE IN A SEPARATE CONTAINER(A CAFETERIA TRAY WILL PROBABLY WORK PERFECTLY).2)
SOFT－AGAR－ASSAY－FOR－COLONY－FORMATION
Note: All volumes are calculated to cater for four plates per point.Base Agar1. Melt 1% Agar (DNA grade) in microwave, cool to 40 in a waterbath. War
In－Vitro－prostate－colony－and－sphereforming－assays
1.?Prostates were dissected, minced into small pieces with a steel blade, and digested with 0.8 mg/ml collagenase in 10 ml of primary cell medium/
造血干細胞CFU（Colony－Forming－Unit）集落形成檢測
實驗概要造血干細胞CFU（Colony Forming Unit）集落形成檢測主要試劑DPBS，HSCs培養液，MethoCult?培養基，HSCs稀釋液主要設備離心機，超凈臺，體視鏡，倒置顯微鏡，35 mm、100 mm培養皿，注射器，16號鈍針頭，移液器，渦旋儀，5 mL、10 mL移液管，離心
Single－tube－confirmation－PCR－protocol
The following colony PCR protocol has been designed to be performed in individual reaction tubes. We usually test three colonies from each transformat
PCR－protocol
PCR reactionProtocol for 50μl reaction - adjust amounts if necessary, for a 20μl reaction use the same volumes of primer and dNTP-mix, but adjust the
Direct－PCR－from－Whole－Yeast－Cells:－Zymolyase－Method
Direct PCR from Whole Yeast Cells: Zymolyase MethodContributor: Namjin ChungDate: June 18, 19961. An average-size yeast colony (0.5-2mm) or a cell pel
DNA克隆
DNA克隆（主要內容如下）·?????????General Procedure·?????????PCR Cloning·?????????Subcloning·?????????ET Cloning·?????????Vector Preparation·?????????Ligation Re
其它PCR方法
·?????????Standard PCR Protocol?(Molecular Biology Techniques Manual)The followings are described in detailRecommended Reagent ConcentrationsRecommend
Streptomyces:Protocols/PCR
Description?Polymerase Chain Reaction (PCR) is a method of amplifying a specific DNA target sequence. The cycle involves denaturing the template doubl
Infusion－biobrick－assembly
OverviewThis is a method to assemble two BioBricks using the Clontech In-Fusion PCR Cloning Kit and maintains BioBrick standard formats. There are cur
實驗室自動化與篩選協會2013亞洲會展新品發布
2 kinds of newly launched products are available to SLAS Exhibitors to distribute on 2013 SLAS Asia Conference & Exhibition website. This is
DNA轉化
DNA轉化Chemical Transformation·?????????Transformation of Competent Cells (RbCl2 Method)?(Goldberg Lab)Very nice protocol for E. Coli transformation inc
Yeast－Gene－knockout－using－Oligo/PCR
Universal primers for gene knock-out using dominant drug markers: Kan, Clonat, and Hygromisin-B.?Forward primer: 5’ TCAGGGGCATGATGTGACT 3’Reverse prim
Troubleshooting－for－PCR－and－multiplex－PCR
Troubleshooting discussion is based on the PCR protocol as described in the table below. All reactions are run for 30 cycles.COMPONENTVOLUMEFINALCONCE
PCR簡介/PCR儀
PCR的要素基本的PCR須具備PCR儀圖冊1.要被復制的DNA模板 Template2.界定復制范圍兩端的引物Primers.3.DNA聚合酶Taq. Polymearse4.合成的原料（四種脫氧核苷酸）及水。 PCR儀工作原理利用升溫使DNA變性，在聚合酶的作用下使單鏈復制成雙鏈，進而達到基因復制
多重PCR（Multiplex－PCR）
一般PCR僅應用一對引物，通過PCR擴增產生一個核酸片段，主要用于單一致病因子等的鑒定.多重PCR(multiplex PCR)，又稱多重引物PCR或復合PCR，它是在同一PCR反應體系里加上二對以上引物，同時擴增出多個核酸片段的PCR反應，其反應原理，反應試劑和操作過程與一般PCR相同.??? 多
多重PCR（Multiplex－PCR）
一般PCR僅應用一對引物，通過PCR擴增產生一個核酸片段，主要用于單一致病因子等的鑒定.多重PCR(multiplex PCR)，又稱多重引物PCR或復合PCR，它是在同一PCR反應體系里加上二對以上引物，同時擴增出多個核酸片段的PCR反應，其反應原理，反應試劑和操作過程與一般PCR相同.??? 多
重疊PCR—overlap－PCR
1、簡介?重疊PCR也是基本的PCR原理：變性-退火-延伸。不同的是在重疊PCR過程是兩個或者幾個片段重疊延伸之后，再進行指數擴增的PCR過程。 ?2、基本原理*步：PCR產生兩個或者幾個片段，這幾個片段之間必須有重疊區。?第二步：以兩個片段為例，見上圖，*步產生的兩個片段，A D鏈之間有互補，B