RNaseandDEPCTreatment:FactorLaboratoryMyth
Researchers are usually trained in RNA isolation and analysis methods by one another or by technical manuals. Experimental procedures are often not questioned and quickly become dogma. Furthermore, it is difficult to find literature to document the "facts" taught by mentors and technical manuals. One of these potential myths is the use of DEPC treatment to make solutions RNase-free. At Ambion, we have syste......閱讀全文
RNase－and－DEPC－Treatment:－Fact－or－Laboratory－Myth
Researchers are usually trained in RNA isolation and analysis methods by one another or by technical manuals. Experimental procedures are often not qu
RNAse－A－Treatment－of－Mouse－Cells
IntroductionRNAse A treatment of permeabilized cells followed by immunostaining is a method which allows to show if the localization of a protein into
DEPC
Procurement100ml DEPC from Sigma (D5758)?[1]?- €233/$292 (as of 2009-01)100ml DEPC from MPI (150902)?[2]?- €313 (as of 2009-01)DEPC-treatment of solut
CIP－Treatment
set up the following reaction:CIP RxnH2O7.8 ml10x cip rxn buffer2.0 mlDNA(e.g; 3 kb vector; 0.2 mg/ml; 2 mg total)10.0 ml(1 u/ml) CIP0.2 mltotal20.0 m
正確使用DEPC
DEPC有致癌性，不能處理Tris溶液等這些事項大家都知道，所以不談。談一些實驗室中的操作誤區。誤區一：重復使用含DEPC的水處理槍頭、離心管等。由于DEPC較貴，而且書上說DEPC要用高壓滅菌處理去除，于是有些人就萌發了重復使用DEPC的想法，即對含有DEPC的水不進行高壓滅菌，從而重復使用含DE
RNase－A/Tl保護和－RNase－H－聚焦法實驗——RNase－H－聚焦法
實驗方法原理RNA與互補的 [?32P ] 標記的探針在溶液中復件后形成的 RNA-RNA 雜合分子對單鏈專一的 RNase 具有抗性。此法可以對 RNA 分子末端進行定位或對含內含子的交界定位。與 Northern blot 相比，它是一種十分有效而且靈敏度很高的并可用于測定 mRNA 豐度的方法
RNase－A/Tl保護和－RNase－H－聚焦法實驗——RNase保護法
基因分子鑒定的一個重要方面就是其 RNA 轉錄物 5' 和 3' 端的精確作圖。首先被 Maniatis 研究組使用的 RNase 保護測定法就是基于這個目的發展起來的。由于雜交是在溶液中實現的，因此也稱為溶液雜交。本實驗來源「RNA 實驗指導手冊」主編：鄭曉飛。實驗方法原理RNA與
MITOMYCIN－C－TREATMENT－OF－PMEFs
Cultures to be treated should be sub confluent ie actively growing.1. Add 1/20 volume Mitomycin C (200 ug/ml 10 ug/ml), to culture and incubate at 37
RNase－A/Tl保護和－RNase－H－聚焦法實驗
RNase保護法 RNase H 聚焦法 ? ? ? ? ? ? 實驗方法原理 RNA與互補的 [ 32P ] 標記的探針在溶液中復件后形成的 RNA-RNA 雜合分子對單
RNase－A/Tl保護和－RNase－H－聚焦法實驗
實驗方法原理 RNA與互補的 [ 32P ] 標記的探針在溶液中復件后形成的 RNA-RNA 雜合分子對單鏈專一的 RNase 具有抗性。此法可以對 RNA 分子末端進行定位或對含內含子的交界定位。與 Northern blot 相比，它是一種十分有效而且靈敏度很高的并可用于測定 mRNA
Formaldehyde－Treatment－of－Tissue－Culture－Hoods
You will need:-12g Potassium Permanganate?6g Crushed paraformaldehyde1) Set the hood so that it can vent outside.2) Mix the two chemicals above togeth
RNase－inhibitors－and－RNases
Question ?1.What are the differences among RNase H, RNase A, RNase B and RNase C?2.In your cDNA kits, RNase H is added in the second strand reaction t
General－Laboratory－Methods－for－Tetrapyrroles
There are thousands of porphyrins and metalloporphyrins, and hundreds of new derivatives appear each year. This variety arises because the cyclic
E.Z.N.A.?－Total－RNA－Maxi－Kit－Protocol－for－Animal－Tissues
實驗概要The E.Z.N.A.? ?Total RNA Maxi Kit uses HiBind? matrix spin-column technology to ?isolate up to 5 mg total cellular RNA from a variety of sources ?
DNAse－posttreatment－for－nuclear－antigens
Rationale:?The use of DNAse to improve nuclear antigen staining has been published long before the AR era?1, 2.DNAse treatment is currently suggested
Oligo(dT)纖維素層析分離Poly(A)+－RNA
Selection of Poly(A)+?RNA by Oligo(dT)-Cellulose ChromatographyJoseph SambrookPeter Maccallum Cancer Institute and The University of Melbourne, Austra
Guidelines－for－the－Use－of－Analgesics－and－Tranquilizers－in－Laboratory－Animal
What is Anesthesia??Anesthesia is a state of unconsciousness induced in an animal. The three components of anesthesia are?analgesia?(pain relief),?amn
An－Integrative－Procedure－for－Apoptosis－Identification－and－Measurement2
TroubleshootingCritical Steps(1) Don’t trypsinize cells for too long when collecting them.(2) Rotation speed should be no more than 1500 rpm during ce
Effect－of－UV－Treatment－on－Early－Development－of－Sea－Urchin－Embryos
Objective:To test the effects of UV radiation at 254nm on Sea Urchin embryo development after radiation at the 2-cell stage.Background:Sea Urchins exh
DEPC水處理的75％酒精如何配制
1. RNase-free玻璃瓶的獲取：用含終濃度0.1%DEPC的超純水（DEPC水）浸泡玻璃瓶過夜，2. 制備RNase-free水：RNase-free玻璃瓶裝上0.1%DEPC的超純水高溫高壓滅菌。3. 75%乙醇（DEPC水配制）：RNase-free-水：無水乙醇（未開封的）按1:3配制
General－Laboratory－Procedures,－Equipment－Use,－and－Safety－Considerations
A. Storage .The following properties of reagents and conditions are important considerations in processing and storing DNA and RNA. Heavy metals promo
Apoptosis:－A－Laboratory－Manual－of－Experimental－Methods－Andrea－Cossarizza
THE CELL?1.?Morphological aspects of apoptosis?Walter Malorni, Stefano Fais & Carla Fiorentini?2.?Cell cycle?Miriam Capri & Daniela BarbieriTHE NUCLEU
Size－and－Shape－of－Protein－Molecules2
The sedimentation coefficient of a protein is a measure of how fast it moves through the gradient. Increasing the mass of the protein will increase it
How－to－make－DEPCtreated－water－and－Tris－Buffer
Add 0.1 ml DEPC to 100 ml of the?solution to be treated and shake vigorously to bring the DEPC into solution.Let the?solution incubate for 12 hours at
RNA抽提注意事項和經驗指南3
RNA 抽提的“三大紀律八項注意” 紀律一：杜絕外源酶的污染。注意一：嚴格戴好口罩，手套。注意二：實驗所涉及的離心管，Tip 頭，移液器桿，電泳槽，實驗臺面等要徹底處理。注意三：實驗所涉及的試劑/溶液，尤其是水，必須確保 RNase-Free。紀律二：阻止內源酶的活性注意四：選擇合適的勻漿方法。注意
Guidelines－for－theUse－of－Analgesics－and－Tranquilizers－in－Laboratory－Animal2
GeneralSome of the drugs listed here do not possess all three criteria for an anesthetic and must be used in combinations to achieve full anesthesia o
Guidelines－for－theUse－of－Analgesics－and－Tranquilizers－in－Laboratory－Animal3
BarbituratesThe barbiturates are an acid ring molecule with various ring substitutes that imbue the drug with different properties. Barbiturates are a
Guidelines－for－theUse－of－Analgesics－and－Tranquilizers－in－Laboratory－Animal4
Other?AnalgesicsAnalgesics are pain relievers most often given after a surgery. Narcotic analgesics have already been described above. Nonsteroidal an
Guidelines－for－theUse－of－Analgesics－and－Tranquilizers－in－Laboratory－Animal5
Anesthetic Induction and MaintenanceInjectable AnesthesiaAnesthetic induction using injectable anesthetics is fairly simple. It involves admininsistra
Guidelines－for－theUse－of－Analgesics－and－Tranquilizers－in－Laboratory－Animal6
?Anesthetic?MachineThe best method of delivering an inhalant anesthetic is with an anesthetic machine. These machines precisely mix the gas with air o