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What is a crystaline precipitate that I get sometimes (not always) when I precipitate my genomic DNA preps and how can I get rid of it? theories are protein(that was not completely removed durin ...

Phenol-based Method for the Isolation of DNA Fragments from Low-Melting Temperature Agarose Reference: Favre D. 1992. Biotechniques vol. 13 1.Cut out slice containing DNA smallest size possible. 2.E ...

1.Inoculate a couple of 20 ml LB broth in a 125 ml flasks with 0.5 ml of an overnight culture of the recipient strain. 2.Shake at 37℃ until O.D. 600=0.13 - 0.15 (1 - 2 hours). Measure by taking ...

Buffer TfbI pH5.8 500ml1.47g KOAC ...

Day 1 1. Digest DNA for 6 hours (or overnight) BSA 10 mg/ml 0.5 22.65 μl of 10 μg Genomic DNA RnaseA 10mg/ml 0.1 in TE mixed to 7.35 μl cocktail Spermidine 100nM 0.75 per sample 10X enzyme b ...

NOTE: THIS IS FINE FOR SOUTHERNS BUT NOT FOR SCREENING BY PCR. (from Ruixia 7/99 from protocol by Stef Oehen 7/94). 1. Put 1 cm tail in 1.5 ml microcentrifuge tube. 2. Add 500µl Tail Buffer an ...

This protocol originated in Murray and Thompson (1980) and then was modified by Richard Jorgensen and finally published in Wagner et. al 1987. 1.Start with about 10 grams fresh needles. 2.Chop into s ...

载体：携带外源DNA 进入宿主细胞的工具。 一、载体的功能 1.运送外源基因高效转入受体细胞。 2.为外源基因提供复制能力或整合能力。 3.为外源基因的扩增或表达提供条件。 二、载体应具备的条件 1.具有对受体细胞的可转移性。 2.具有与特定受体细胞相适应的复制位点或整合位点。 3.长度尽可能小，以提高其载装能力。 4.具有多种单一的酶切位点。 5.具有合适的选择性标记。 附图： ...

Uniplex PCR-based Sequencing Latest update 2-21-00 A.PCR Reactions: B.Processing PCR products: C.Alternative PCR Reaction Cleanup steps instead of passage through Sephadex G-50 usually not performe ...

Two protocols are given here. The first one is used for BAC library screening on filters although is also suitable for Transfer hybridizations. PREPARATION OF HYBRIDIZATION SOLUTION Prehybridizati ...

Hi there Is anyone know how short of the CpG island can be? let say if the CG sequence of about 30bp is that possible the short CpG sequence be methylated? Thanks you! Pine -pine- ------------------- ...

Saghai-Maroof MA Soliman KM Jorgensen RA & Allard RW (1984) PNAS 81:8014-8018 DNA successfully isolated from fungal species of Cochliobolus Aternaria and Fusarium. The key elements in this ...

Phenol-chloroform DNA extraction from sand flies 1. Crash Individual sand flies in 0.5 ml lysis buffer (50 mM NaCl 10 mM EDTA 50 mM Tris-HCl pH 8). 2. Incubate The sand fly homogena ...

Michael Koelle's LabDepartment of Molecular Biophysics and Biochemistry Yale University http://info.med.yale.edu/mbb/koelle/protocols/protocol_subcloning.html ...

Map-Based Cloning of KAM2 Map-based cloning of the KAM2 gene was performed essentially as described previously (Tamura et al. 2005). We used codominant cleaved amplified polymorphic sequence markers ( ...

These procedures were originally devised in Richard Palmiter's lab for use with tail dots (DNA spotted onto a nitrocellulose filter and probed for a transgene; quicker than doing southerns) and subseq ...

macerate tissue in Eppendorf tube without butter at RT add 400 ml extraction buffer vortex for 4 sec leave sample at RT until other samples are ready (> 1 h) spin in microfuge for 1 min ...

To efficiently determine the chromosomal location of phenotypic mutants we designed a genome-wide mapping strategy that can be used in any crop for which a dense AFLP (Amplified Fragment Length Polymo ...

Author: Laura-Lee Boodram Source: Laura-Lee Boodram Department of Life Sciences The University of the West Indies Abstract: The protocol is simple and fairly rapid. It does not require the use of orga ...

自杀性质粒载体完全可以自己构建，只要保证两点： 1.自杀性质粒载体不能在你的宿主菌里进行复制，也就是说载体上不能有能在宿主菌中复制的复制子； 2.要由筛选标记． 自杀性质粒载体不能在你的宿主菌里进行复制，否则会导致细菌染色体突变株的构建失败。但是在构建时，要注意以下方面： 1、在制备质粒载体和DNA插入片断时 2μg 5kb大小的线状DNA大约含有1.4poml/L5'末端磷酸。5&lsqu ...