The present invention discloses a high-GC content of PCR gene amplification buffer. 所述高GC含量基因的PCR扩增缓冲液包括如下组分:TrisHcl150mmol/L,(NH4)2SO440mmol/L,一水甜菜碱2.6mol/L,乳化剂Tween 20 0.02%,PCR反应增强剂20%,配置后,过滤去菌,20℃保存. The GC-rich PCR gene amplification ...
我做的是链霉菌,所以GC含量也在70多。我P的效果很好 5.可以试着加点DMSO、NaOH、甘油等东西试试。以解除其二级结构的影响,当然现在也有专门这种PCR Buffer卖. 6. 如果实在不行,就用advantage 2 high GC taq polymeras.(clontech),效果好,就是贵了点。 可以用双温法试试,比如35个循环,总变性之后,前五个...
1、PCR中GC含量过高的问题DNA molecule: 4849Length = 1212 base pairsMolecular Weight = .00 Daltons, single strandedMolecular Weight = .00 Daltons, double strandedG+C content = 70.96%A+T content = 29.04%Nucleotide Number Mol% A 173 14.27 C 438 36.14 G 422 34.82T 179 14.77解决:1.由于基因...
DNAsegmentswithveryhighGCcontenthaveproveddifficulttohandleinawiderangeofmolecularanalyses.GC-richregionsmayformrigid,constrainedsecondarystructuresthataredifficultorimpossiblefortheDNApolymerasestoenterunderstandardPCRconditions.6 HighlyGC-richregionscanobstructPCRdependentanalysesinthefollowingsituations:1.StandardPCR...
2. Melting TemperaturePrimers with similar Tm, preferably between 55°C-60°C are used. For sequences with high GC content, primers with a higher Tm (preferably 75°C-80°C) are recommended. A Tm variation of between 3°-5° C is acceptable for primers used in a pool. ...
GC buffer 效果很好 我做的是链霉菌 所以 GC 含量也在 70 多我 P 的效果很好 5 可以试着加点 DMSO NaOH 甘油等东西试试 以解除其二级结构的影响 当然现在也有专门这种 PCR Buffer 卖 6 如果实在不行 就用 advantage 2 high GC taq polymeras clontech 效果好 就是贵了点 可以用双温法试试 比如 35 ...
In the present study we optimized the PCR conditions for the amplification of the human ADAMTS-2 gene promoter region featuring an extremely high GC content and a secondary structure. We show that a combination of three additives, betaine, dimethyl sulfoxide, and 7-deaza GTP, is essential to ...
False-negative and false-positive results can be generated by NGS-based platforms in genomic regions with highly repetitive sequence, high GC content, or homologous sequences. The regions with high GC content or repetitive sequences are often underrepresented in the NGS coverage because of bias in...
Zhang and collaborators recently described a TCN coctail which in combination with inhibitor-resistant Taq DNA polymerase mutants enabled efficient amplification of high-GC content DNA targets directly from crude blood samples [27]. However, when wild-type Taq DNA polymerase was used for PCR with TC...
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