ColonyHybridization
ProcedurePrepare serial 10-fold dilutions of transformed bacteria in LB and spread 100 μL onto LB/Amp plates, as described in the Electroporation protocol. Incubate at 37°C overnight. Also streak out positive and negative control bacteria on a separate plate.Select plates that have an optimal density of bacteria (i.e. 2 mm spacing between colonies). Put selected plates at 4°C for one hour.Lay 83 mm circular nylon mem......閱讀全文
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)
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－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
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
In－situ－hybridization
Note:?Although it is possible to perform in situ hybridization on bleached embryos, it appears to reduced the strength of the signal. For best results
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
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
Bacterial－Colony－PCR
Bacterial Colony?PCRObjective:This protocol allows rapid detection of transformation success when primers are available to allow determination of corr
Microarray－Hybridization－Protocol
Introduction:One microarray set consists of 7 nylon membranes with 2.5 x 7.5 cm dimension. 2304 genes were spotted onto nylon membranes (Schleicher an
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
ORNL－MICROARRAY－HYBRIDIZATION－PROTOCOLS
Direct labeling of total RNA with Cy3 and Cy5:A. MATERIALSRNeasy? Mini Kit (Qiagen; Cat # 74106)?SuperScript II RT (200U/μL) (Life Technologies; Cat #
Principles－of－nucleic－acid－hybridization
Principles of nucleic acid hybridization5.2.1.?Nucleic acid hybridization is a method for identifying closely related nucleic acid molecules within tw
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
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
Basic－Fluorescent－in－situ－Hybridization－(FISH)
實驗概要Fluorescence ?in situ hybridization method is a kind of physical map drawing method, ?use fluorescent element mark probe, to detect probe and spli
In－Situ－Hybridization－to－Somatic－Chromosomes－in－Drosophila
In Situ Hybridization to Somatic Chromosomes in DrosophilaAbby F. DernburgINTRODUCTIONIn situ hybridization was originally developed as a technique fo
Making－RNA－probes－for－in－situ－hybridization
Make DNA templates via PCRDay 1It's preferable to start with constructs that contain RNA polymerase start sites (T3, T7, or SP6) which allow use o
Fluorescence－In－Situ－Hybridization－using－TSA?
實驗概要This ?protocol describes steps for fluorescent in situ hybridization (FISH) ?to Drosophila embryos using Tyramide Signal Amplification (TSA?), and
CDNA文庫
?CDNA文庫（主要內容如下）·?????????Construction of cDNA Library·?????????Construction of Genome DNA Library·?????????Library Screening??OthersConstruction of cD
Screening－BAC－filters－with－nonradioactive－probes
(Protocol for a single high-density BAC colony filter (HDR filter) of 22 cmx22 cm) to be screened using Amersham''s ECL hybridization kit; thi
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
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/
Engineering－BioBrick－vectors－from－BioBrick－parts/Colony－PCR
MaterialsPCR SuperMix High FidelityVF2 primer (5''-TGCCACCTGACGTCTAAGAA-3'')VR primer (5''-ATTACCGCCTTTGAGTGAGC-3'')De
熒光原位雜交（Fluorescence－in－situ－hybridization,FISH）
實驗原理熒光原位雜交（Fluorescence in situ hybridization FISH）是一門新興的分子細胞遺傳學技術，是20世紀80年代末期在原有的放射性原位雜交技術的基礎上發展起來的一種非放射性原位雜交技術。目前這項技術已經廣泛應用于動植物基因組結構研究、染色體精細結構變異分析、病
Hybridization－of－High－Density－Arrayed－BAC－Nylon－Filter－Blots
Protocol for hybridization of high density arrayed Bacterial Artificial Chromosome nylon filter blots with 100 PCR isolated Unigene cDNA inserts, pool
DNA轉化
DNA轉化Chemical Transformation·?????????Transformation of Competent Cells (RbCl2 Method)?(Goldberg Lab)Very nice protocol for E. Coli transformation inc
熒光原位雜交（Fluorescence－in－situ－hybridization,FISH）（圖）
實驗原理熒光原位雜交（Fluorescence in situ hybridization FISH）是一門新興的分子細胞遺傳學技術，是20世紀80年代末期在原有的放射性原位雜交技術的基礎上發展起來的一種非放射性原位雜交技術。目前這項技術已經廣泛應用于動植物基因組結構研究、染色體精細結構變異分析、病
熒光原位雜交（Fluorescence－in－situ－hybridization,FISH）原理
2）標本變性①將制備好的染色體玻片標本于 50oC培養箱中烤片2～3h。（經Giemsa染色的標本需預先在固定液中退色后再烤片）。②取出玻片標本，將其浸在70～75oC的體積分數70％甲酰胺/2×SSC的變性液中變性2～3min。③立即按順序將標本經體積分數70％、體積分數90％和體積分數100％冰
雙重和多重原位雜交(hybridization－in－situ)技術
為了在同一標本上或同一細胞內同時檢測是否存在兩種或兩種以上的靶核酸序列。可應用雙重或多重原位雜交技術．即以兩種或多種標記探針與靶核酸雜交。然后利用不同的檢測手段分別顯示各種靶核酸的存在和分布。該技術與免疫組織化學技術中的雙重或多重標記相似，除了探針本身的特異性外，對結果的干擾主要來自標記物及檢測試劑