PEGPRECIPITATIONOFPCRPRODUCTS
PEG PRECIPITATION OF PCR PRODUCTSThis protocol can be used instead of EXO/SAP for removing excess primers and nucleotides from PCR products before cycle-sequencing.Protocol:1. After PCR, add the following reagents to each tube:For 30 μl reaction add:4.8 μl of 5 M NaCl 4.8 μl of TE buffer 8.4 μl of 40% PEG-8000, 10mM MgCl2For 50 μl reaction a......閱讀全文
PEG-PRECIPITATION-OF-PCR-PRODUCTS
PEG PRECIPITATION OF PCR PRODUCTSThis protocol can be used instead of EXO/SAP for removing excess primers and nucleotides from PCR products before cyc
Ethanol-precipitation-method-for-purifying-PCR-products
1. For each PCR reaction (50 μL) prepare a 1.5mL microcentrifuge tube containing the following:??????? - 5 μL of 3M sodium acetate (NaOAc), pH 4.6????
Ethanol-precipitation-method-for-purifying-PCR-products
1. For each PCR reaction (50 μL) prepare a 1.5mL microcentrifuge tube containing the following:??????? - 5 μL of 3M sodium acetate (NaOAc), pH 4.6????
Isolation-Of-PCR-Products
實驗概要Rapid and efficient purification of PCR products from salts, primers, dNTPs, and other non-nucleic acid reagents.實驗原理The ChargeSwitch? TechnologyT
Electrophoresis-of-PCR-products-with-Sunrise-gel-apparatus
Electrophoresis of PCR products with Life Technologies Sunrise gel apparatusGel:?In a 500 ml Pyrex? glass bottle, add:Agarose:3 gH2O270 mls10X TA30 ml
Cloning-PCR-products-using-TA-vectors
Cloning PCR products using TA vectorsby Paul N. Hengen, Ph.D.?*Methods and reagents is a unique monthly column that highlights current discussions in
High-Throughput-Isolation-Of-PCR-Products-Using-ChargeSwitch?-PCR-CleanUp
實驗概要The ChargeSwitch? ?PCR Clean-Up Kit allows rapid and efficient purification of PCR ?products from salts, primers, dNTPs, and other non-nucleic aci
Template-Preparation
Template PreparationThe quality of sequencing results is directlyrelated to the quality of the template.?ABI recommends a minialkaline-lysis/PEG preci
CORE-SAMPLE-PCR:-A-method-to-rePCR-unique-bands-from-products-of-mixed-s
INTRODUCTIONThe products of a PCR reaction - especially when this is done on eukaryotic genomic DNA, and when using degenerate primers - often contain
Acetone-precipitation-of-protein
This procedure is suitable for recovering proteins from most aqueous solvents and from SDS containing buffers. It is not recommended for proteins diss
TRFLP技術的優缺點
T-RFLP(末端限制性片段長度多態性)該技術在應用的過程中,肯定需要在實驗條件上不斷進行改進,而這種改進的好壞自然而然需要實驗結果的驗證。V. Grüntzig在2002年做了該工作的一部分,結果認為,在限制性酶切時,很有必要去除其中影響DNA的酶切過程,并且實驗證明了具體的酶切時間。具有說服力的
TRFLP的優缺點
該技術在應用的過程中,肯定需要在實驗條件上不斷進行改進,而這種改進的好壞自然而然需要實驗結果的驗證。。V. Grüntzig在2002年做了該工作的一部分,結果認為,在限制性酶切時,很有必要去除其中影響DNA的酶切過程,并且實驗證明了具體的酶切時間。具有說服力的結果如下:??1,T-RFLP出數據的
DNA抽提
DNA抽提(主要內容如下)·???Working with DNA·???DNA Extraction from Bacteria and Other Organisms·???DNA Extraction from Cell and Tissue·???Mitochondria DNA Isola
PCR的下游應用
·?????????Agarose Gel Electrophoresis of PCR Products?(Robert H. Cruickshank)·?????????Agarose Gel Electrophoresis of PCR Products?(Immunology Resourc
PCR的下游應用
??????????Agarose Gel Electrophoresis of?PCR?Products(Robert H. Cruickshank)??????????Agarose Gel Electrophoresis of PCR Products(Immunology Resource)
PCR基本實驗方法(四)
?Labelling?PCR?Products with DigoxigeninPCR products may be very conveniently labelled with digoxigenin-11-dUTP (Boehringer-Mannheim) by incorporating
PCR基本實驗方法(四)
Labelling?PCR Products with DigoxigeninPCR products may be very conveniently labelled with digoxigenin-11-dUTP (Boehringer-Mannheim) by incorporating
基于PCR技術的染色質沉淀分析2
METHOD?Analysis of precipitated chromatin fractions (from?Chromatin Immunoprecipitation on Unfixed Chromatin from Cells and Tissues to Analyze Histone
Protocol-for-Trichloroacetic-Acid-(TCA)-Precipitation
IntroductionThe efficiency of nucleotide incorporation in DNA/RNA polymerization reactions (e.g. transcription, reverse transcription, and DNA replica
DNA-methyltransferase-Assay
Methylated CpG island Amplification?Protocol written by Minoru Toyota2. Materials2.1. MCARestriction enzymes SmaI, XmaIT4 DNA ligaseTaq DNA polymerase
PIC-Crosslinking-and-Immune-Precipitation
References?Fishburn et. al., 2005, Molecular Cell, vol. 18 #3: Experimental Procedures pg. 376Immobilized Template assay (Hahn lab website)NotesDTT mu
沉淀反應技術(Precipitation-reaction-technique)
一、???? 概述?可溶性抗原(如細菌浸出液、含菌病料浸出液、血清以及其他來源的蛋白質、多糖質、類脂體等)與其相應的抗體相遇后,在電解質參與下,抗原抗體結合形成白色絮狀沉淀,出現白色沉淀線,此種現象稱為沉淀反應。沉淀反應中的抗原叫沉淀原(precipitinogen),與沉淀原發生反應的抗體稱為沉淀
PEG是什么
1、PEG是聚乙二醇英文名polyethylene glycol的縮寫,是一種化學藥品,被廣泛的應用于化妝品行業。化學結構由環氧乙烷聚合而成。液體PEG采用200Kg鍍鋅桶包裝;膏狀PEG采用50Kg廣口桶包裝;固體PEG采用25Kg塑料襯里紙板桶或編織袋包裝。該品無毒、難燃,可按一般化學品運輸規定
沉淀反應技術(Precipitation-reaction-technique)(1)
一、 概述可溶性抗原(如細菌浸出液、含菌病料浸出液、血清以及其他來源的蛋白質、多糖質、類脂體等)與其相應的抗體相遇后,在電解質參與下,抗原抗體結合形成白色絮狀沉淀,出現白色沉淀線,此種現象稱為沉淀反應。沉淀反應中的抗原叫沉淀原(precipitinogen),與沉淀原發生反應的抗體稱為沉淀素(pre
沉淀反應技術(Precipitation-reaction-technique)(2)
2.取試管5支(5mm×50mm)置于試管架上,編號。第1、2試管內加炭疽沉淀血清,第3、4試管內加正常血清,第5管內加待檢抗原,分別用毛細滴管加至4mm~5mm。3.第1、4、5試管輕輕疊加等量緩沖液,第2、3試管輕輕疊加等量待檢抗原。為防止上下兩界面破壞,可將小試管從試管架取出,微傾斜,沿試管壁
Analysis-of-Intermediates-and-End-Products-of-the-Chlorophyll-Bio...
Analysis of Intermediates and End Products of the Chlorophyll Biosynthetic PathwaySince the 1963 seminal review of Smith and French (62), our unders
蛋白產物(protein-products)的檢測實驗
一、組織蛋白的裂解提取:1.分別取-70℃保存的各組動物的樣品(半個腦、0.5g肌肉),放入小離心管中,在冰浴上以PBS充分洗滌2次,除去殘留血液。2.用無菌手術剪和鑷子將大鼠肌肉剪碎(腦樣品不用剪碎),放入另一小離心管中,于0℃以 PBS充分洗滌,4℃,3000 rpm離心5分鐘。3.吸出上清夜,
DNA測序
DNA測序(主要內容如下)·?????????Sequencing Gel Preparation·?????????Preparation of Templates?·?????????DNA Sequencing by the Dideoxy Method·?????????DNA Sequen
Taq酶PCR實驗方法介紹
General AdvicePCR allows the production of more than 10 million copies of a target DNA sequence from only a few molecules. The sensitivity of this tec
M13噬菌體
·?????????M13 Phage?(Michael Blaber)Very useful background information about M13: its infection, replication, packing, cloning. If you are new to phag