Reagents & EquipmentsOligo-dT (T15) - Roche dNTPs (in buffer 50 mM Tris pH 8) RNasin - Promega Superscript II - Life Technologies Qiaquick PCR purification system - QIAGEN 22 x 50 mm cover slips Cot-1 ...
IntroductionChromatin immunoprecipitation coupled to tiling microarray analysis (ChIP-on-chip) is used to measure genome-wide the DNA binding sites of a protein of interest. In ChIP-on-chip proteins a ...
IntroductionHybridization of chromatin immuno-precipitation (ChIP) material to tiling arrays at NimbleGen service facilities usually leaves the customer with a set of data files that are of limited us ...
IntroductionChromatin immunoprecipitation (ChIP) arguably represents the most powerful application of antibody technology to epigenetic research. It allows analysis of patterns of histone modification ...
Materials for 30 arrays Qty Order info Humid chamber 1 Sigma #H 6644 Inverted heat block (70-80C) 1 Diamond scriber 1 VWR #52865-005 Slide rack 1 Shandon Lipshaw #121 Slide chamber 2 Shandon Lipshaw ...
One rxn One 96-well plate (100 rxns) RESGEN SPECIFIC PRIMERS: Program: 92C (30") 56C (45") 72C (3'30") / 36 cycles Forward primer (20 uM) 5 - Reverse primer (20 uM) 5 - 10X PCR buffer (Perkin Elmer) 1 ...
Note: for single spotted arrays halve volumesMix two color samples (for genomic DNA Normally 0.5ug) add 2ml Block Mix (10mg/ml salmon sperm DNA (BRL) + 4mg/ml yeast tRNA)Use Speed Vec to dry the sampl ...
RNA preparation We recommend the following protocols for extracting total RNA: cultured cells: Quiagen RNeasy Kit Trizol (Gibco) tissues: Quiagen RNeasy Kit ...
Materials Qty Order info Glass microscope slides 60 Gold Seal #3010 Slide rack 2 Shandon Lipshaw #121 (800-245-6212) Slide chamber 6 Shandon Lipshaw #121 ddH2O ~5 L NaOH 70 g 95% Ethanol 420 mL Poly-L ...
MaterialsRNA sample 30 µg Total or 2 µg mRNAOligo dT (dT18VN) 5 µg/µL 34.8 (µg/mL)/A260Superscript II 5x 1st strand buffer (Invitrogen)1 M DTT (Sigma 43816 $16.80/10 mL)50x dNTP Mix (dATP dCTP dGTP @2 ...
The following is a slight modificatioundefined of a protocol developed by Joe DeRisi (UCSF) and Rosetta Inpharmatics (Kirkland WA). Original document can be obtained at www.microarrays.org.A. RT Reaction ...
Resuspend the contents of one pack of the monofunctional NHS-ester Cy3 or Cy5 dye (mono-functional Cy3 or Cy5 reactive pack Amersham catalog PA 23001 and PA 25001) in 72 ul DMSO. Aliquot 16 x 4.5 ul a ...
I. Preparing fluoresenctly labeled cDNA (probe):To anneal primer mix 2ug of mRNA or 50-100mg total RNA with 4ug of a regular or anchored oligo-dT primer in a total volume of 15.4 ul: Cy3 Cy5 mRNA (1 ...
Materials for 2 reactions (Cy3 & Cy5) Qty Order info Heat blocks 42C & 70C 1 each Oligo dT (2 ug/uL) 5.0 uL Superscript II RT kit Gibco BRL #18064-014 50X dNTPs (25 mM; 10 mM dTTP~undefined 1.25 uL Cy3-dUTP ( ...
目录第一章总RNA的纯化（RNeasy®Kit）.............................3第二章由RNA合成双链DNA..............................................4一、反转录合成第一链cDNA............................................4二、反转录合成第二链cDNA... ...
Easy Way to Clone The protocol is oriented towards a C. albicans genomic library I made in Lambda Zap II on 7/97. The overall sequence of events is: • Titer and plate out phage • Lift plaques onto fil ...
Cosmid Cloning: Cell preparation DNA packaging and Cell TransfectionProtocol taken from Stratagene's Gigapack packaging extracts instruction manual Host bacteria (E. coli LE392)preparation:Pick one co ...
Day OneStart overnight cultures of Cosmid in fresh L-Broth + 25 ug/ml kanamycin. 4 x 250 ml each inoculate each with metal loop from frozen glycerol stocks. Grow cultures for 12 hours(do not let go fo ...
DNA Isolation by a Cleared Lysate Method Followed by Double Acetate Precipitation Version 3b - updated September 26 1999 The Most Recent Roe Lab Implementation (by Feng Chen Hau-Qin Pan and Fu Ying) A ...
Plasmid or Cosmid DNA MiniprepThis protocol can be used to isolate sufficient amount DNA from 1.5ml o/n culture or 3ml 6hr culture to do several enzyme digestions.Spin 1.5ml o/n culture at 12000rpm fo ...
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