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Digest DNA in 96-well plateTo each well add:4ul 10Xbuffer4ul Enzyme0.4ul Spermidine(0.4M)31.6ul H2O37?C 19h, then add 4ul loading dye to each well. Load into 400ml 1% agarose gel immediately or keep the plate at -20?C.Southern Blot1) Take picture of agarose gel to be blotted with phoshorescent ruler lined up along side it, such that the ruler is lined up with the top of the wells. This is so you can later estimate th......閱讀全文
哈嘍，小伙伴們！走過不要錯過，又到了科普知識的時間了，喜歡做分子實驗的小伙伴一定聽說過Southern blot這門檢測技術，但大家知道為什么叫Southern嗎？今天小編就為大家科普一下這門實驗技術。 Southern Blot是最早出現的blot（印跡）技術，它是檢測DNA的一種方法。
Picking ES cell clonesOne or two days before picking colonies prepare 24-well plates of feeders. You can also use alternate protocols that utilize 96-
圖4. Southern blot鑒定同源重組事件[4]另外，在制備基因敲進和條件性基因敲除模式小鼠過程中，Southern blot檢測是非常重要的一步。在我們以往的工作中，其中一例Cre定點敲進課題（如圖5），Southern blot檢測結果顯示：小鼠1號，有明顯的隨機插入現象。圖5
DNA PreparationGene TransferEmbryo TransferTransgenic IdentificatioinOthersTransgenic Outline (University of Michigan Transgenic Animal Model Cor
Need 1.5-2 x 107 cells from a 2 day culture.1. Cells are harvested as normal, washed x 1 in PBS then taken up at conc. of 1.2 x 10 7 cells/ml in cold
HYBRIDIZATION.Prehybridize blot at 65oC for ~3h in Church buffer containing 0.5mg/ml denature salmon sperm DNA (usually 14ml Church buffer plus 0.7ml
實驗概要DNAzol? Reagent (Genomic DNA Isolation Reagent) is a complete and ready-to-use reagent for the isolation of genomic DNA from solid and
YWB per 10ml5mL 2M Sorbitol (if NZ arrested, add 40uL 1.5mg/mL0.336mL 1M K2HPO4 N2 to 4mL YWB)0.064mL 1M KH2PO44.6mL dH2OYWB, glycerol, PMSF5mL 2
Koshland Lab,Carnegie Institute http://www.ciwemb.edu/labs/koshland/Protocols/MICROTUBULE/mmb.htmlDetermine the OD600 and correlate the cell
Methylated CpG island Amplification Protocol written by Minoru Toyota2. Materials2.1. MCARestriction enzymes SmaI, XmaIT4 DNA ligaseTaq DNA polym
Megabase DNA IsolationMegabase-size DNA isolation from plantsTo construct large insert DNA libraries in BAC and YAC vectors, methods must be developed
實驗概要The E.Z.N.A.? Blood DNA Maxi Kit is designed for isolation of genomic DNA from up to 25 ml of fresh, whole blood treated with any comm
NUCLEAR EXTRACTION OF TISSUE CELLSThis procedure was developed for HNEK cells grown in KGM, from Clonetics. All steps are performed on ice, with
Preparation of nucleic acid probesIn standard nucleic acid hybridization assays the probe is labeled in some way. Nucleic acid probes may be made as s
實驗概要The E.Z.N.A.? Blood DNA Maxi Kit is designed for isolation of genomic DNA from up to 25 ml of fresh, whole blood treated with any comm
Differential cDNA Screening ProceduresThe protocols listed refer to cDNA library construction and preliminary differential screening procedures. They
Cell cycle analysis of Escherichia coli cellsC period = the time for a round of chromosome replicationD period = the time between the end of
AbstractWhen more than one bait will be used to screen a single library, significant time and resources can be saved by performing the interactor hunt
實驗概要stem cell culture protocol主要試劑cell culture supplies and reagentssEnvironment: cell culture requires a sterile environment, so it needs a separat
ContentsEmbryonic Stem Cell MarkersHematopoietic Stem Cell MarkersMesenchymal/Stromal Stem Cell MarkersNeural Stem Cell MarkersReferencesWhile stem ce
co-IP assays can be performed between endogenous proteins or transiently or stably expressed exogenous - usually tagged - proteins. The advantage to u
We use feeder-independent ES cell lines derived from the 129/Ola strain of mice (Nichols et al., Development 110, p.1341, 1990). These cells are easy
MediaHigh glucose DMEM (-pyruvate, -glutamine)20% Heat-inactivated Fetal calf serum (can vary by cell type, be sure!!)1X l-glutamine1X Penicillin/stre
哈嘍，小伙伴們！走過不要錯過，又到了科普知識的時間了，喜歡做分子實驗的小伙伴一定聽說過Southern blot這門檢測技術，但大家知道為什么叫Southern嗎？今天小編就為大家科普一下這門實驗技術。 Southern Blot是最早出現的blot（印跡）技術，它是檢測DNA的一種方法。
· Standard PCR Protocol (Molecular Biology Techniques Manual)The followings are described in
Southern雜交One important thing for transfer:the weight of the object resting on top of the blotting apparatus should not exceed the weight equal to a 5
PREPARE SOLUTIONS1. 10mM MgSO4, 0.2% Maltose LB (100 mL):Mix 1.0 g of Bacto-Tryptone, 1.0 g of NaCl, 0.5 g of Yeast Extr
Mammalian RNA InterferenceThomas TuschlLaboratory for RNA Molecular BiologyThe Rockefeller University, New York Excerpted from RNAi: A Guide
實驗概要The PureLink? Genomic DNA Purification Kit allows rapid and efficient purification of genomic DNA. The kit is designed to efficiently
人胚腎 293 (HEK293) 細胞在重組蛋白表達中是最常見的宿主細胞。 這類細胞能夠表達大量的膜蛋白，如 G 蛋白偶聯受體 (GPCR) ，是無法在最常見的生物制藥生產宿主，如：中國倉鼠卵巢 (CHO) 細胞中作表達。 HEK293 雖然是蛋白表達的極好宿主，然而 H