PCR引物设计

Plant MaterialsCotton ovules (Gossypium hirsutum cv. Coker 312) were collected 8 15 and 20 days after anthesis. Total RNA was extracted from stripped ovules (fibers removed) and isolated fibers using ...

MaterialsROX Passive Reference Dye 25 µM 50x (Invitrogen Cat. No. 12223-012)SYBR Green I "10000x" concentrate (Molecular Probes Cat. No. S7563)SureStart Taq polymerase 5 U/µL (Stratagene Cat. No. 6002 ...

Mutant allele: Neo-1 primer (CCTTCTATCGCCTTCTTG) plus mgK-3 primer (TGGAACCCTGTGCCATCTCTAT) produce a 0.5kB PCR product.Wildtype allele: mgK-3 (above) plus mcK-5 (GAGCAGACGCCCAAGAAGC) produce ~1kB pro ...

一、引物设计step by step1、在NCBI上搜索到目的基因，找到该基因的mRNA，在CDS选项中，找到编码区所在位置，在下面的origin中，Copy该编码序列作为软件查询序列的候选对象。2、用Primer Premier5搜索引物①打开Primer Premier5，点击File-New-DNA sequence 出现输入序列窗口，Copy目的序列在输入框内（选择As），此窗口内，序列也 ...

PCR reactionProtocol for 50µl reaction - adjust amounts if necessary for a 20µl reaction use the same volumes of primer and dNTP-mix but adjust the volume of water and 10x buffer for a 100µl reaction ...

For each 20 ul of PCR product prepare the following mixture in 1.5 ml Tube. 2 ul of 3M sodium acetate (NaOAc) pH 4.5 40 ul chilled absolute ethanol. Transfer the PCR product (20ul) into the tube conta ...

Initial denaturation It is very important to denature the template DNA completely. Initial heating of the PCR mixture for 2 minutes at 94°�95°C is enough to completely denature complex genomic DNA so ...

A variety of PCR additives and enhancing agents have been used to increase the yield specificity and consistency of PCR reactions. Whilst these additives may have beneficial effects on some amplificat ...

Since PCR is often problematic on GC-rich templates we have evaluated different PCR enhancing additives and generated an Combinatorial Enhancer Solution (CES). We tested this solution on approx. 50 di ...

Degenerate PCR is in most respects identical to ordinary PCR but with one major difference. Instead of using specific PCR primers with a given sequence you use mixed PCR primers. That is if you do not ...

Isolate partially purified chromosomal DNA using a modification of the Magic MiniPreps DNA Purification System from Promega (see atta ched) as follows. - Spin down 500 μl of an overnight culture in a ...

Following PCR you often want to get rid of the PCR primers and Taq polymerase before the next step. This is necessary for sequencing PCR products or TA cloning. There is no clean up needed following g ...

Multiplex_protocol_NP2006.pdf

1) Immunocapture stageCoating buffer: 15 mM Na2CO3; 35 mM NaHCO3 and 3 mM NaN3 per liter (pH 9.6). Extraction buffer: (20 mM Tris-HCL (pH 8.0) 138 mM NaCl 1 mM PVP 0.05% Tween-20 3 mM KCl and 3 mM Na ...

DescriptionSingle primer PCR allows amplification from known to unknown regions in chromosomes phage plasmids large PCR products and other sources of DNA.At sufficiently low stringency any primer will ...

Colony PCR is useful in determining whether or not a specific colony on a plate has a sequence you desire. Primers for the specific sequence should be used when preparing the reaction cocktail. Exampl ...

TAIL is a series of reactions that are intended to map where a T-DNA (transfer DNA) has inserted within the genome. The main components of the 3 reactions are the AD (Arbitrary Degenerate) primers bor ...

RT-PCR or reverse transcription PCR refers to PCR that uses product of an RT reaction as template. In effect the PCR amplifies cDNA fragments. In one-tube RT-PCR RNA and PCR primers are added to a rea ...

RNA Extraction from Histologic SectionsUnstained 4 µm thick sections of formalin fixed paraffin embedded liver biopsies were transferred from glass slides to 1.5 ml non-siliconized microcentrifuge tub ...

Cut PVDF membrane to the appropriate size activate with absolute methanol for 5 sec and incubate in distilled water for 5 min. For electroblotting equilibrate in transfer buffer and follow the standar ...